TW202227464A - Multiepitope vaccine for the treatment of alzheimer’s disease - Google Patents

Abstract

The disclosure provides peptide compositions and immunotherapy compositions comprising an amyloid-beta (A[beta]) peptide and an alpha-synuclein peptide. The disclosure also provides methods of treating or effecting prophylaxis of Alzheimer's disease or other diseases with beta-amyloid deposition in a subject, including methods of clearing deposits, inhibiting or reducing aggregation of A[beta] and/or alpha-synuclein, blocking the uptake by neurons, clearing amyloid, and inhibiting propagation of alpha-synuclein seeds in a subject having or at risk of developing Alzheimer's disease or other diseases containing alpha-synuclein and/or amyloid-beta accumulations. The methods include administering to such patients the compositions comprising an amyloid-beta (A[beta]) peptide and an alpha-synuclein peptide.

Description

Multi-epitope vaccine for Alzheimer's disease

The present invention relates to the technical fields of immunology and medicine, and in particular to the treatment of Alzheimer's disease and other diseases of protein misfolding.

Alzheimer's disease (AD) is a progressive disease that causes dementia in the elderly. In general, the disease is divided into two categories: late-onset disease, which occurs in old age (over 65 years of age); and early-onset disease, which develops before old age, that is, between the ages of 35 and 60. to some extent. The pathology is the same in both types of disease, but tends to produce more severe and extensive abnormalities in cases that begin at a younger age. The disease is characterized by at least two types of lesions in the brain, neurofibrillary tangles and aging plaques. Aging plaques (ie, amyloid plaques) are regions of chaotic neural fiber networks up to 150 μm long interspersed with extracellular amyloid deposits in the center, which can be detected by microscopic analysis of brain tissue sections. The accumulation of amyloid-like plaques in the central nervous system is also associated with Down's syndrome and other cognitive disorders (cerebrovascular amyloid (CAA)) and eye disease (age-related macular degeneration).

The major components of plaques are peptides called A[beta] or [beta]-amyloid peptides. A[beta] peptides are 4-kDa internal fragments of 38 to 43 amino acids of a large transmembrane glycoprotein called amyloid precursor protein (APP). As a result of the proteolytic processing of APP by different secretases, A[beta] exists mainly in the short form (40 amino acids in length) and the long form (ranging from 42 to 43 amino acids in length). A portion of the hydrophobic transmembrane domain of APP is present at the carboxy-terminus of A[beta] and may explain the ability of A[beta] to aggregate into plaques, especially in the long form. The accumulation of amyloid plaques in the brain eventually causes neuronal cell death. Alzheimer's disease can be characterized by cognitive and physical symptoms associated with this type of neurodegeneration.

Alpha-synuclein is a protein present in neurons and other cells that is a major component of the pathology that characterizes several neurodegenerative disorders, collectively referred to as synuclein disorders, including Parkinson's Parkinson's disease, dementia with Lewy body and multiple system atrophy. The normal physiological function of alpha-synuclein is limited, but there is evidence that the soluble form of the protein can interact with other proteins and certain intracellular membranes. In synuclein disorders, alpha-synuclein appears to accumulate abnormally within cells, which is the pathology of the disease. There is increasing evidence that certain aggregated forms of alpha-synuclein can be transported from neuron to neuron, causing the spread of pathology that results in neuronal dysfunction and loss. In some neurodegenerative diseases, misfolding and aggregation of α-synuclein (SNCA) is often accompanied by β-amyloid deposition, and α-synuclein and Aβ aggregates coexist in some neurodegenerative diseases Diseases include Alzheimer's disease and Parkinson's disease.

Accordingly, there is a need for novel therapies and agents for the prevention or treatment of Alzheimer's disease, particularly those capable of eliciting an immune response against Aβ and α-synuclein present in patients.

In some embodiments, the invention relates to polypeptides comprising a first peptide comprising 3 to 10 amino acids from residues 1 to 10 of SEQ ID NO: 01 linked to a second peptide, the The second peptide includes 3 to 10 amino acids from residues 81 to 140 of SEQ ID NO:02. For example, the second peptide can be from the C-terminus of alpha-synuclein (residues 111 to 140 of SEQ ID NO:02). The first peptide can be N-terminal to the second peptide, or the first peptide can be C-terminal to the second peptide. Furthermore, the first peptide can include the amino acid sequence of one of SEQ ID NOs: 3 to 38 or 121 to 176, and the second peptide can include the amino acid sequence of one of SEQ ID NOs: 39 to 109 . For example, the first polypeptide can be DAEFRHD (SEQ ID NO:06), DAEFR (SEQ ID NO:08) or EFRHD (SEQ ID NO:21), and the second polypeptide can be 5 to 10 amine groups Acids such as PDNEAYE (SEQ ID NO:55) or DPDNEAYE (SEQ ID NO:48).

In other embodiments, the first peptide and the second peptide can be linked by a cleavable linker, which can be an amino acid sequence. The cleavable peptide linker, if present, can be 1 to 10 amino acids in length. In some embodiments, the linker comprises about 1 to 10 amino acids, about 1 to 9 amino acids, about 1 to 8 amino acids, about 1 to 7 amino acids, about 1 to 6 amino acids amino acid, about 1 to 5 amino acids, about 1 to 4 amino acids, about 1 to 3 amino acids, about 2 amino acids, or one (1) amino acid. In some embodiments, the cleavable peptide linker is 1 amino acid, 2 amino acids, 3 amino acids, 4 amino acids, 5 amino acids, 6 amino acids, 7 amino acids Amino Acids, 8 Amino Acids, 9 Amino Acids, or 10 Amino Acids. For example, the linker can be arginine-arginine (Arg-Arg), arginine-valine-arginine-arginine (Arg-Val-Arg-Arg (SEQ ID NO: 113 )), valine-citrulline (Val-Cit), valine-arginine (Val-Arg), valine-lysine (Val-Lys), valine-alanine (Val) -Ala), phenylalanine-lysine (Phe-Lys), glycine-alanine-glycine-alanine (Gly-Ala-Gly-Ala; SEQ ID NO: 114), Ala-Gly-Ala -Gly (SEQ ID NO: 115) or Lys-Gly-Lys-Gly (SEQ ID NO: 116). In particular embodiments, the polypeptide can be DAEFRHDRRPDNEAYEGGC (SEQ ID NO: 110) or DAEFRHDRRDPDNEAYEGGC (SEQ ID NO: 111).

In other embodiments, the polypeptide may include a linker at the C-terminal portion of the polypeptide or the N-terminal portion of the polypeptide to the vector. The linker, if present, can be 1 to 10 amino acids in length. In some embodiments, the linker comprises about 1 to 10 amino acids, about 1 to 9 amino acids, about 1 to 8 amino acids, about 1 to 7 amino acids, about 1 to 6 amino acids amino acid, about 1 to 5 amino acids, about 1 to 4 amino acids, about 1 to 3 amino acids, about 2 amino acids, or one (1) amino acid. In some embodiments, the linker is 1 amino acid, 2 amino acids, 3 amino acids, 4 amino acids, 5 amino acids, 6 amino acids, 7 amino acids , 8 amino acids, 9 amino acids, or 10 amino acids. For example, a linker can include the amino acid sequences of GG, GGG, AA, AAA, KK, KKK, SS, and SSS. Additionally, if present at the C-terminus, the linker attached to the carrier may include a C-terminal cysteine (C). Alternatively, if present at the N-terminus, the linker attached to the carrier may include an N-terminal cysteine (C). For example, a polypeptide can include the amino acid sequence of DAEFRHDRRX ₁ PDNEAYEXXC (SEQ ID NO: 112), wherein X ₁ is optional, and if present, X ₁ is D, and wherein XX and C are independently optional Conditions exist, and if present, XX can be GG, AA, KK, SS, GAGA (SEQ ID NO: 114), AGAG (SEQ ID NO: 115), or KGKG (SEQ ID NO: 116).

In other embodiments, the invention pertains to an immunotherapy composition comprising a polypeptide of the invention, wherein the polypeptide can be linked to a carrier. Carriers may include serum albumin, immunoglobulin molecules, thyroglobulin, ovalbumin, tetanus toxoid (TT), diphtheria toxoid (DT), genetically modified cross-reactive substance (CRM) of diphtheria toxin, CRM197, meninges Pneumococcal outer membrane protein complex (OMPC) and Haemophilus influenzae protein D ( H. influenzae protein D ; HiD), rEPA (Pseudomonas aeruginosa exotoxin A), KLH (keyhole snail) limpet) hemocyanin) and flagellin.

Furthermore, embodiments of the present invention pertain to a pharmaceutical formulation comprising a polypeptide or immunotherapy composition of the present invention and comprising at least one adjuvant. The adjuvant can be aluminum hydroxide, aluminum phosphate, aluminum sulfate, 3-de-O-phosphorylated monophosphoryl lipid A (MPL), QS-21, QS-18, QS-17, QS-7, TQL1055, Complete Freund's Adjuvant (CFA), Incomplete Freund's Adjuvant (IFA), oil-in-water emulsions (such as squalene or peanut oil), CpG, polyglutamic acid, polylysine, AddaVax™ , MF59® and combinations thereof. Additionally, formulations may include liposomal formulations, diluents, or multiple antigen presentation systems (MAPs). MAPs can include Lys-based dendritic architectures, helper T cell epitopes, immunostimulatory lipophilic moieties, cell penetrating peptides, free radical-induced polymerization, self-assembling nanoparticles (as an antigen presentation platform), and Chennai One or more of the rice particles.

Furthermore, embodiments of the present invention relate to immunotherapy compositions comprising a first peptide sequence comprising 3 to 10 amines from the first ten N-terminal residues of SEQ ID NO:01 and a second peptide sequence amino acid residues, and the second peptide sequence comprises 3-8 amino acids from residues 81 to 140 of SEQ ID NO:02. The first peptide can include the amino acid sequence of one of SEQ ID NOs: 3-38 or 121-176, and the second peptide can include the amino acid sequence of one of SEQ ID NOs: 39-109. Each of the first peptide and the second peptide can include a linker at the C-terminal portion of the polypeptide to the carrier. The linker, if present, can include an amino acid sequence selected from the group consisting of GG, AA, KK, SS, GAGA (SEQ ID NO: 114), AGAG (SEQ ID NO: 115), and KGKG (SEQ ID NO: 116 ), and may include a C-terminal cysteine (C). Carriers may include serum albumin, immunoglobulin molecules, thyroglobulin, ovalbumin, tetanus toxoid (TT), diphtheria toxoid (DT), genetically modified cross-reactive substance (CRM) of diphtheria toxin, CRM197, meninges Pneumococcal outer membrane protein complex (OMPC) and Haemophilus influenzae protein D (HiD), rEPA (Pseudomonas aeruginosa exotoxin A), KLH (keyhole hemocyanin) and flagellin.

In addition, the immunotherapy composition can include at least one pharmaceutically acceptable diluent and/or a multiple antigen presentation system (MAP). MAPs can include Lys-based dendritic architectures, helper T cell epitopes, immunostimulatory lipophilic moieties, cell penetrating peptides, free radical-induced polymerization, self-assembling nanoparticles (as an antigen presentation platform), and Chennai One or more of the rice particles.

The immunotherapy composition can be included in a pharmaceutical composition comprising the immunotherapy composition and at least one adjuvant, such as aluminum hydroxide, aluminum phosphate, aluminum sulfate, 3-de-O-acylated monophosphoryl Lipid A (MPL), QS-21, TQL1055, QS-18, QS-17, QS-7, complete Freund's adjuvant (CFA), incomplete Freund's adjuvant (IFA), oil-in-water emulsions such as squalene or peanut oil), CpG, polyglutamic acid, polylysine, AddaVax™, MF59®, and combinations thereof.

Embodiments of the invention also pertain to nucleic acid sequences encoding the polypeptides and immunotherapy compositions of the invention. Nucleic acids can be included in nucleic acid immunotherapy compositions comprising the nucleic acid and at least one adjuvant.

Furthermore, embodiments of the present invention relate to methods of treating or preventing Alzheimer's disease in an individual, and inhibiting or reducing Alzheimer's disease in individuals suffering from or at risk of developing Alzheimer's disease Methods of aggregation of at least one of A[beta] and [alpha]-synuclein. The methods include administering to an individual an immunotherapy composition, nucleic acid immunotherapy composition or pharmaceutical formulation of the invention.

The methods of the present invention may include at least two, at least three, at least four, at least five, or at least six repeated administrations, and may include repeated administrations at intervals of about 21 to about 28 days.

Furthermore, the methods of the present invention are directed to inducing immune responses in animals. The method comprises administering to the animal a polypeptide, immunotherapy composition, pharmaceutical formulation or nucleic acid immunotherapy composition of the invention in a regimen effective to generate an immune response comprising specific binding to Aβ, α-synuclein or Antibodies to both A[beta] and [alpha]-synuclein. The immune response may include antibodies that specifically bind to the N-terminal region of A[beta] and/or the C-terminal region of [alpha]-synuclein.

In other embodiments, the invention pertains to an immunization kit comprising an immunotherapy composition of the invention and can include an adjuvant, wherein the immunotherapy composition can be in a first container and the adjuvant can be in a second container middle.

Furthermore, the present invention relates to a kit comprising the nucleic acid immunotherapy composition of the present invention and may include an adjuvant. The nucleic acid can be in a first container and the adjuvant can be in a second container.

related applications

This application claims the rights of US Provisional Application No. 63/080,619, filed September 18, 2020, which is incorporated herein by reference in its entirety.

The present invention provides peptide compositions and immunotherapy compositions comprising amyloid-β (Aβ) peptides and α-synuclein peptides. The present invention also provides methods of treating or achieving prevention of Alzheimer's disease or other diseases with β-amyloid deposition in a subject, including in patients with Alzheimer's disease or other diseases containing α-synuclein and Clearing and preventing deposit formation in diseases of or at risk of developing amyloid-beta accumulation, inhibiting or reducing Abeta and/or alpha-synuclein aggregation, blocking binding by neurons and/or uptake of A[beta] and/or alpha-synuclein, inhibiting the transport of alpha-synuclein species between cells, and inhibiting the spread of pathology between brain regions. The method includes administering to such patients a composition comprising an amyloid-beta (Abeta) peptide and an alpha-synuclein peptide.

The following are definitions of various terms. As used herein, the singular forms "a/an" and "the (the)" include plural references unless the context clearly dictates otherwise. For example, the terms "compound" or "at least one compound" can include plural compounds, including mixtures thereof.

Unless otherwise apparent from the context, the term "about" encompasses insubstantial variations, such as values within standard measurement error margins (eg, SEM) of the stated values. For example, when referring to a measurable value such as a parameter, amount, duration, the term "about" as used herein can encompass a difference of +/- 10% or less, +/- 5% from the specified value % or less, or +/- 1% or less or less. A specification of a range of values includes all integers within or bounding that range, and all subranges bounded by integers within that range. As used herein, statistical significance means p≤0.05.

A composition or method that "comprises" or "includes" one or more of the recited elements can include other elements not specifically recited. For example, a composition that "comprises" or "includes" a polypeptide sequence may contain the sequence alone or in combination with other sequences or components.

If an individual has at least one known risk factor (eg, age, genetics, biochemistry, family history, and contextual exposure), the individual is at increased risk of developing the disease, and the risk of developing the disease in individuals with that risk factor is statistically significant were significantly greater than individuals who did not have this risk factor.

The term "patient" includes human and other mammalian individuals receiving prophylactic or therapeutic treatment, including untreated individuals. As used herein, the term "subject" or "patient" refers to any single subject in need of treatment, including other mammalian subjects, such as humans, cows, dogs, guinea pigs, rabbits, and the like. An individual is also intended to include any individual participating in a clinical research trial that does not exhibit any clinical signs of disease, or participating in an epidemiological study, or serving as a control.

The term "disease" refers to any abnormal condition that impairs physiological function. The term is used broadly to encompass any disorder, disease, abnormality, disorder, disease, condition or syndrome that impairs physiological function, regardless of the nature of the cause.

The term "symptom" refers to subjective evidence of a disease as perceived by an individual, such as a change in gait. "Symptoms" means objective evidence of disease as observed by a physician.

As used herein, the term "treat/treatment" refers to alleviating or alleviating one or more symptoms or effects associated with a disease; preventing, inhibiting or delaying the onset of one or more symptoms or effects of a disease; reducing the The severity or incidence of one or more symptoms or effects; and/or increase or cause a desired outcome as described herein.

As used herein, the term "prevention/prevent/preventing" refers to the presence or absence of Aβ and/or α-synuclein pathology (primary and secondary) prior to the onset of disease prophylaxis), contacting (e.g., administering) a peptide or immunotherapy composition of the invention to an individual, thereby delaying the onset of clinical symptoms and/or relieving symptoms of the disease after the onset of the disease (with the individual not being combined with the peptide or immunotherapy) compared to exposure to physical substances), and does not imply complete suppression of disease onset. In some cases, prophylactic effects may be maintained for a limited time after administration of the peptides or immunotherapy compositions of the invention. In other cases, prophylaxis may be maintained for the duration of a treatment regimen comprising administration of the peptides or immunotherapy compositions of the invention.

As used herein, the term "reduction/reduce/reducing" refers to reducing or suppressing an increase in the amount of Aβ and/or α-synuclein present in an individual or a tissue of an individual, which encompasses reducing or An increase in the amount of Aβ and/or α-synuclein present, accumulated, aggregated or deposited in an individual or a tissue of an individual is inhibited (eg, the rate of increase is decreased). In certain embodiments, reducing or inhibiting an increase in the amount of Aβ and/or α-synuclein present, accumulated, aggregated or deposited in an individual (eg, reducing the rate of increase) refers to a central nervous system in the individual The amount of A[beta] and/or [alpha]-synuclein present, accumulated, aggregated or deposited in the system (CNS). In certain embodiments, reducing or inhibiting an increase in the amount of Aβ and/or α-synuclein present, accumulated, aggregated, or deposited in an individual (eg, reducing the rate of increase) refers to a peripheral ( For example, the amount of A[beta] and/or [alpha]-synuclein present, accumulated, aggregated or deposited in the peripheral circulatory system. In certain embodiments, reducing or inhibiting an increase in the amount of Aβ and/or α-synuclein present, accumulated, aggregated, or deposited in an individual (eg, by reducing the rate of increase) means that in the individual's brain The amount of Aβ and/or α-synuclein present in, accumulated, aggregated or deposited in. In some embodiments, the reduced A[beta] and/or alpha-synuclein is a pathological form of: A[beta] (eg, beta-amyloid (A[beta]) extracellular plaque deposits, neuroinflammatory amyloid plaques block) and/or α-synuclein (eg, fibular-like α-synuclein inclusions, oligomeric or fibrillar α-synuclein coalescing, and α-synuclein oligomer precursors fiber intermediates). In other embodiments, pathological indicators of neurodegenerative diseases and/or synuclein disorders are reduced.

The term "epitope" or "antigenic determinant" refers to a site on an antigen to which B cells and/or T cells can respond, or a site on an antigen to which an antibody binds. Epitopes can be formed from adjacent amino acids or non-adjacent amino acids, which are juxtaposed by tertiary folding of the protein. Epitopes formed from adjacent amino acids typically remain after exposure to denaturing solvents, whereas epitopes formed by tertiary folding typically disappear after treatment with denaturing solvents. Epitopes typically include at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12 in unique spatial configurations or at least 13 amino acids. Methods for determining the spatial configuration of epitopes include, for example, x-ray crystallography and 2-dimensional nuclear magnetic resonance. See, eg, Epitope Mapping Protocols, Methods in Molecular Biology, Vol. 66, Ed. Glenn E. Morris (1996).

An "immunogenic agent" or "immunogen" or "antigen" is capable of inducing an immune response against itself or against a modified/treated version of itself upon administration to an animal, optionally combined with an adjuvant. The term "immunogenic agent" or "immunogen" or "antigen" refers to a compound or composition comprising a peptide, polypeptide or protein which, when administered in an appropriate amount (an "immunogenically effective amount"), Polypeptides or proteins are "antigenic" or "immunogenic", that is, capable of inducing, eliciting, increasing or enhancing a cellular and/or humoral immune response and are recognized by the products (T cells, antibodies) of that response. The immunogen can be a peptide or a combination of two or more identical or different peptides including at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, or at least 13 amino acids in a linear or steric configuration.

An immunogen can be effective when provided alone or in combination, or linked or fused to another substance, which can be administered all at once or at several intervals. An immunogenic agent or immunogen can include an antigenic peptide or polypeptide linked to a carrier as described herein.

A nucleic acid, such as DNA or RNA, that encodes an antigenic peptide or polypeptide is referred to as a "DNA [or RNA] immunogen" because the encoded peptide or polypeptide is expressed in vivo following administration of the DNA or RNA. A peptide or polypeptide can be expressed recombinantly by a vaccine vector, which can be naked DNA or RNA, comprising a peptide or polypeptide coding sequence operably linked to a promoter (eg, an expression vector or cassette as described herein) .

The term "adjuvant" refers to a compound that enhances an immune response to an antigen when administered in combination with an antigen, but does not generate an immune response to the antigen when administered alone. Adjuvants can enhance the immune response by several mechanisms, including lymphocyte recruitment, stimulation of B cells and/or T cells, and stimulation of macrophages. Adjuvants can be natural compounds, modified versions or derivatives of natural compounds, or synthetic compounds.

The terms "peptide" and "polypeptide" are used interchangeably herein and refer to a chain of two or more consecutive amino acids. If and when a distinction is made, the context should make the meaning clear and unambiguous. For example, if two or more peptides described herein are joined to make a dimeric or multimeric peptide, then polypeptides can be used to designate "multiple" or "more than one" peptides.

The term "pharmaceutically acceptable" means that the carrier, diluent, excipient, adjuvant or adjuvant is compatible with the other ingredients of the pharmaceutical formulation and is not substantially harmful to its receptor.

The term "immunotherapy" or "immune response" refers to the production of beneficial humoral (antibody-mediated) and/or cellular (antigen-specific T cells) directed against Aβ and/or α-synuclein peptides in the recipient. or its secreted product mediated) response. Such responses may be active responses induced by administration of an immunogen, such as A[beta] and/or [alpha]-synuclein peptides. Cellular immune responses are elicited by the presentation of polypeptide epitopes in conjunction with MHC class I or class II molecules to activate antigen-specific CD4 ⁺ T helper cells and/or CD8 ⁺ cytotoxic T cells. Responses may also involve activation of monocytes, macrophages, NK cells, basophils, dendritic cells, astrocytes, microglia, eosinophils, or other components of innate immunity. The presence of cell-mediated immune responses can be determined by proliferation assays (CD4 ⁺ T cells) or CTL (cytotoxic T lymphocytes) assays. The relative contributions of humoral and cellular responses to the protective or therapeutic effect of an immunogen can be distinguished by isolating antibodies and T cells individually from immunized isogenic animals and measuring the protective or therapeutic effect in a second individual.

beta amyloid ( A beta or Abeta )

A[beta] (also referred to herein as amyloid beta peptide or Abeta) peptide is an approximately 4-kDa internal fragment of APP with 38 to 43 amino acids (A[beta]39, A[beta]40, A[beta]41, A[beta]42 and A[beta]43). For example, A[beta]40 consists of residues 672 to 711 of APP and A[beta]42 consists of residues 673 to 713 of APP. As a result of the proteolytic processing of APP in vivo or in situ by different secretases, Aβ is predominantly in the "short form" (40 amino acids in length) and the "long form" (ranging in length from 42 to 43 amino acid) is present. As described herein, the epitope or epitope is located within the N-terminus of the Aβ peptide and includes within amino acids 1 to 10 and 12 to 25 of Aβ, eg, residues 1 to 3, 1 to 4 from Aβ , 1 to 5, 1 to 6, 1 to 7, or 3 to 7 of Aβ, or 2 to 4, 2 to 5, 2 to 6, 2 to 7, or 2 to 8, residues 3 to 5, 3 of Aβ to 6, 3 to 7, 3 to 8 or 3 to 9, or residues 4 to 7, 4 to 8, 4 to 9 or 4 to 10 of Aβ, residues 12 to 24, 12 to 23, 12 to 10 of Aβ 22, 13 to 25, 13 to 24, 13 to 23, 13 to 22, 14 to 25, 14 to 24, 14 to 23, 14 to 22, 15 to 25, 15 to 24, 15 to 23, or 15 to 22. For example, residues 12-17, 12-18, 12-19, 12-20, 12-21, 13-17, 13-18, 13-19, 13-20, 13-21, 13 from Aβ42 to 22, 14 to 17, 14 to 18, 14 to 19, 14 to 20, 14 to 21, 14 to 22, 14 to 23, 15 to 17, 15 to 18, 15 to 19, 15 to 20, 15 to 21 , 15 to 22, 15 to 23 or 15 to 24. Other examples of epitopes or epitopes include residues 16-18, 16-19, 16-20, 16-21, 16-22, 16-23, 16-24, 16-25, 17-19 of Aβ42 , 17 to 20, 17 to 21, 17 to 22, 17 to 23, 17 to 24 or 17 to 25. Other examples of epitopes or epitopes include residues 18-20, 18-21, 18-22, 18-23, 18-24, 18-25, 19-21, 19-22, 19-23 of Aβ42 , 19 to 24, 19 to 25, 20 to 22, 20 to 23, 20 to 24, 20 to 25, 21 to 23, 21 to 24 or 21 to 25.

Aβ (Abeta) is a major component of the plaques characteristic of Alzheimer's disease. A[beta] is produced by the processing of the larger protein APP by two enzymes, termed beta and gamma secretase. Mutations in APP known to be associated with Alzheimer's disease occur near sites of beta-secretase or gamma-secretase or within A[beta]. A portion of the hydrophobic transmembrane domain of APP is present at the carboxy-terminus of A[beta] and may account for the ability of A[beta] to aggregate into plaques, especially if it is elongated. The accumulation of amyloid plaques in the brain eventually causes neuronal cell death. Alzheimer's disease is characterized by physical symptoms associated with this type of neurodegeneration.

alpha - synuclein

Alpha-synuclein is a highly conserved protein that is abundant in neurons, especially at the front end of the synapse. Aggregated alpha-synuclein forming brain lesions is a hallmark of neurodegenerative synuclein disorders. In addition, in some neurodegenerative diseases, misfolding and aggregation can often be accompanied by β-amyloid deposition, and α-synuclein is present in several neurodegenerative disorders, including Alzheimer's and Parkinson's disease.

of immunogens Aβ / alpha - synuclein polypeptide

Agents for active immunization can induce an immune response in patients and can act as immunotherapy. The agent used for active immunization can be, for example, the same type of immunogen used to generate monoclonal antibodies in experimental animals, and can include 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 or more adjacent amino acids.

In some embodiments of the invention, the Aβ/α-synuclein immunogen may comprise an Aβ peptide linked to an α-synuclein peptide comprising the N-terminal sequence from Aβ (SEQ ID NO: 01 ) from 3 to 10 amino acids of residues 1 to 10 of α-synuclein peptide comprising 3 to 10 of residues 81 to 140 of α-synuclein (SEQ ID NO:02) amino acid. For example, an alpha-synuclein peptide may comprise 3 to 10 amino acids from the C-terminal region of alpha-synuclein (residues 111 to 140 of SEQ ID NO: 02). In some embodiments, the peptide is unphosphorylated. In some embodiments, the peptide is phosphorylated at serine (S), threonine (T), and/or tyrosine (Y) phosphorylation sites.

In some embodiments of the invention, the A[beta] peptide may comprise 3 to 10 amino acids from residues 1 to 10 or 12 to 25 of DAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVVIA (SEQ ID NO: 01). For example, the Aβ peptide is selected from the following: DAEFRHDSGY (SEQ ID NO:03), DAEFRHDSG (SEQ ID NO:04), DAEFRHDS (SEQ ID NO:05), DAEFRHD (SEQ ID NO:06), DAEFRH (SEQ ID NO:07), DAEFR (SEQ ID NO:08), DAEF (SEQ ID NO:09), DAE (SEQ ID NO: 10), AEFRHDSGY (SEQ ID NO: 11), AEFRHDSG (SEQ ID NO: 12), AEFRHDS (SEQ ID NO: 13), AEFRHD (SEQ ID NO: 14), AEFRH (SEQ ID NO: 15), AEFR (SEQ ID NO: 16), AEF (SEQ ID NO: 17), EFRHDSGY (SEQ ID NO: 18), EFRHDSG (SEQ ID NO: 19), EFRHDS (SEQ ID NO:20), EFRHD (SEQ ID NO: 21), EFRH (SEQ ID NO: 22), EFR (SEQ ID NO:23), FRHDSGY (SEQ ID NO: 24), FRHDSG (SEQ ID NO: 25), FRHDS (SEQ ID NO: 26), FRHD (SEQ ID NO: 27), FRH (SEQ ID NO:28), RHDSGY (SEQ ID NO:29), RHDSG (SEQ ID NO:30), RHDS (SEQ ID NO:31), RHD (SEQ ID NO:32), HDSGY (SEQ ID NO: 33), HDSG (SEQ ID NO:34), HDS (SEQ ID NO:35), DSGY (SEQ ID NO:36), DSG (SEQ ID NO:37), SGY (SEQ ID NO:38), VHHQKLVFFA (SEQ ID NO: 121), VHHQKLVFF (SEQ ID NO: 122), VHHQKLVF (SEQ ID NO: 123), VHHQKLV (SEQ ID NO: 124), VHHQKL (SEQ ID NO: 125), HHQKLVFFAE (SEQ ID NO: 126), HHQKLVFFA (SEQ ID NO: 127), HHQKLVFF (SEQ ID NO: 128), HHQKLVF (SEQ ID NO: 129), HHQKLV (SEQ ID NO: 130), HHQKL (SEQ ID NO: 131), HQKLVFFAED (SEQ ID NO: 132), HQKLVFFAE (SEQ ID NO: 133), HQKLVFFA (SEQ ID NO: 134), HQKLVFF (SEQ ID NO: 135), HQKLVF (SEQ ID NO: 136), HQKLV (SEQ ID NO: 137), HQKL (SEQ ID NO: 138), QKLVFFAEDV (SEQ ID NO: 139), QKLVFFAED (SEQ ID NO: 140), QKLVFFAE (SEQ ID NO: 141), QKLVFFA (SEQ ID NO: 142), QKLVFF (SEQ ID NO: 143), QKLVF (SEQ ID NO: 144), QKLV (SEQ ID NO: 145), QKL (SEQ ID NO: 146), KLVFFAEDVG (SEQ ID NO: 147), KLVFFAEDV (SEQ ID NO: 148), KLVFFAED (SEQ ID NO: 149), KLVFFAE (SEQ ID NO: 150), KLVFFA (SEQ ID NO: 151), KLVFF (SEQ ID NO: 152), KLVF (SEQ ID NO: 153), KLV (SEQ ID NO: 154), LVFFAEDVG (SEQ ID NO: 155), LVFFAEDV (SEQ ID NO: 156), LVFFAED (SEQ ID NO: 157), LVFFAE (SEQ ID NO: 158), LVFFA (SEQ ID NO: 159), LVFF (SEQ ID NO: 160), LVF (SEQ ID NO: 161), VFFAEDVG (SEQ ID NO: 162), VFFAEDV (SEQ ID NO: 163), VFFAED (SEQ ID NO: 164), VFFAE (SEQ ID NO: 165), VFFA (SEQ ID NO: 166), VFF (SEQ ID NO: 167), FFAEDVG (SEQ ID NO: 168), FFAEDV (SEQ ID NO: 169), FFAED (SEQ ID NO: 170), FFAE (SEQ ID NO: 171), FFA (SEQ ID NO: 172), FAEDVG (SEQ ID NO: 173), FAEDV (SEQ ID NO: 174), FAED (SEQ ID NO: 175), and FAE (SEQ ID NO: 176).

In certain embodiments, the A[beta] peptide is DAEFRHD (SEQ ID NO:06), DAEFR (SEQ ID NO:08) or EFRHD (SEQ ID NO:21).

The alpha-synuclein peptide may correspond to a peptide comprising 3 to 10 amino acids from residues 81 to 140 of SEQ ID NO:02. In some embodiments, alpha-synuclein is unphosphorylated. In some embodiments, alpha-synuclein is phosphorylated. In some compositions, the alpha-synuclein peptide is selected from the group consisting of: VDPDNEAYEM (SEQ ID NO:39), VDPDNEAYE (SEQ ID NO:40), VDPDNEAY (SEQ ID NO:41), VDPDNEA (SEQ ID NO:42), VDPDNE (SEQ ID NO:43), VDPDN (SEQ ID NO:44), VDPD (SEQ ID NO:45), VDP (SEQ ID NO:46), DPDNEAYEM (SEQ ID NO:47), DPDNEAYE (SEQ ID NO:48), DPDNEAY (SEQ ID NO:49), DPDNEA (SEQ ID NO:50), DPDNE (SEQ ID NO:51), DPDN (SEQ ID NO:52), DPD (SEQ ID NO:53), PDNEAYEM (SEQ ID NO:54), PDNEAYE (SEQ ID NO:55), PDNEAY (SEQ ID NO:56), PDNEA (SEQ ID NO:57), PDNE (SEQ ID NO:58), PDN (SEQ ID NO:59), DNEAYEM (SEQ ID NO:60), DNEAYE (SEQ ID NO:61), DNEAY (SEQ ID NO:62), DNEA (SEQ ID NO:63), DNE (SEQ ID NO:64), NEAYEM (SEQ ID NO:65), NEAYE (SEQ ID NO:66), NEAY (SEQ ID NO:67), NEA (SEQ ID NO:68), EAYEM (SEQ ID NO:69), EAYE (SEQ ID NO:70), EAY (SEQ ID NO:71), AYEM (SEQ ID NO:72), AYE (SEQ ID NO:73), YEM (SEQ ID NO:74), ATGFVKKDQL (SEQ ID NO:75), ATGFVKKDQ (SEQ ID NO:76), ATGFVKKD (SEQ ID NO:77), ATGFVKK (SEQ ID NO:78), ATGFVK (SEQ ID NO:79), ATGFV (SEQ ID NO:80), ATGF (SEQ ID NO:81), ATG (SEQ ID NO:82), TGFVKKDQL (SEQ ID NO:83), TGFVKKDQ (SEQ ID NO: 84), TGFVKKD (SEQ ID NO:85), TGFVKK (SEQ ID NO:86), TGFVK (SEQ ID NO:87), TGFV (SEQ ID NO:88), TGF (SEQ ID NO:89), GFVKKDQL (SEQ ID NO:90), GFVKKDQ (SEQ ID NO:91), GFVKKD (SEQ ID NO:92), GFVKK (SEQ ID NO: 93), GFVK (SEQ ID NO:94), GFV (SEQ ID NO:95), FVKKDQL (SEQ ID NO:96), FVKKDQ (SEQ ID NO:97), FVKKD (SEQ ID NO:98) FVKK (SEQ ID NO: 99), FVK (SEQ ID NO: 100), VKKDQL (SEQ ID NO: 101), VKKDQ (SEQ ID NO: 102), VKKD (SEQ ID NO: 103), VKK (SEQ ID NO: 104), KKDQL (SEQ ID NO: 105), KKDQ (SEQ ID NO: 106), KKD (SEQ ID NO: 107), KDQL (SEQ ID NO: 108), and KDQ (SEQ ID NO: 109).

In some embodiments, the Aβ and/or α-synuclein peptides are linked to form a dual Aβ/α-synuclein polypeptide. The A[beta] and [alpha]-synuclein peptides can be linked by an intrapeptide linker. For example, a polypeptide linker is located between the C-terminus of the first peptide and the N-terminus of the second peptide. The A[beta] peptide and/or the alpha-synuclein peptide can be located in a dual A[beta]/[alpha]-synuclein polypeptide in any order, with or without the intrapeptide linker. For example, the Aβ peptide can be located in the N-terminal portion of the dual polypeptide, and the α-synuclein peptide can be located in the C-terminal portion of the dual polypeptide. Alternatively, the alpha-synuclein peptide can be located on the N-terminal portion of the dual polypeptide, and the Aβ peptide can be located on the C-terminal portion of the dual polypeptide side of the alpha-synuclein peptide. References herein to the first peptide or the second peptide are not intended to indicate the order of the A[beta] and/or [alpha]-synuclein peptides in the polypeptide of the immunogen.

In addition, the C-terminal portion of the A[beta] peptide, alpha-synuclein or dual A[beta]-[alpha]-synuclein polypeptide may include a linker for binding the peptide or polypeptide to the carrier. Linkers that couple the peptide or dual polypeptide to the carrier can include, for example, GG, GGG, KK, KKK, AA, AAA, SS, SSS, GAGA (SEQ ID NO: 114), AGAG between the peptide or dual polypeptide and the carrier (SEQ ID NO: 115), KGKG (SEQ ID NO: 116), and analogs thereof, and may further include a C-terminal or N-terminal cysteine to provide short peptide linkers (eg, G-G-C-, K-K-C-, A-A-C- or S-S-C-). In some embodiments, the linker comprises AA, AAA, KK, KKK, SS, SSS, AGAG (SEQ ID NO: 115), GG, GGG, GAGA (SEQ ID NO: 114), and KGKG (SEQ ID NO: 116 ) amino acid sequence of any one. In some embodiments, any of the Aβ peptide, α-synuclein peptide, and dual Aβ/α-synuclein polypeptide can include a C-terminal cysteine in the absence of a spacer, such as AEFRHDSGC (SEQ ID NO: 117) and DAEFRHDC (SEQ ID NO: 118). In some embodiments, any of the Aβ peptide, the α-synuclein peptide, and the dual Aβ/α-synuclein polypeptide can include an N-terminal cysteine in the absence of a spacer.

When the Aβ and α-synuclein polypeptides are linked to form a dual Aβ/α-synuclein polypeptide, the linker can be a cleavable linker. As used herein, the term "cleavable linker" refers to any linker between antigenic peptides that facilitates or otherwise renders Aβ peptides and α-synuclein peptides less cleavable than those without such cleavable linkages Equivalent peptides of the progeny are more easily separated from each other by cleavage (eg, by endopeptidase, protease, low pH, or any other method that can be performed in or around the antigen-presenting cell) and are thereby more easily separated from each other. Antigen presenting cell processing. In some compositions, the cleavable linker is a protease-sensitive dipeptide or oligopeptide cleavable linker. In certain embodiments, the cleavable linker is sensitive to cleavage by a protease in the trypsin family of proteases. In some compositions, the cleavable linker comprises an amino acid sequence selected from the group consisting of: arginine-arginine (Arg-Arg), arginine-valine-arginine-spermine acid (Arg-Val-Arg-Arg; SEQ ID NO: 113), valine-citrulline (Val-Cit), valine-arginine (Val-Arg), valine-lysine (Val-Lys), valine-alanine (Val-Ala), phenylalanine-lysine (Phe-Lys), glycine-alanine-glycine-alanine (Gly-Ala-Gly- Ala; GAGA (SEQ ID NO: 114)), alanine-glycine-alanine-glycine (Ala-Gly-Ala-Gly; AGAG (SEQ ID NO: 115)), and lysine-glycine Acid-Lysine-Glycine (Lys-Gly-Lys-Gly; KGKG (SEQ ID NO: 116)). In some compositions, the cleavable linker is arginine (Arg-Arg).

In some embodiments of the invention, the dual Aβ/α-synuclein polypeptide comprises DAEFRHDRRPDNEAYEGGC (SEQ ID NO: 110) or DAEFRHDRRDPDNEAYEGGC (SEQ ID NO: 111) or DAEFRHDRRX ₁ PDNEAYEXXC (SEQ ID NO: 112) amino acid sequence, wherein _X is optionally present, and if present, it is D, and wherein XX and C are independently optionally present, and if present, XX can be GG, AA, KK, SS , GAGA (SEQ ID NO: 114), AGAG (SEQ ID NO: 115) or KGKG (SEQ ID NO: 116).

In some embodiments, the dual Aβ/α-synuclein polypeptide is as follows: [ first peptide ]-[ linker 1]-[ second peptide ]-[ linker 2]-[Cys] , where if [ the first peptide] is an Aβ peptide, then [the second peptide] is an α-synuclein peptide, and if [the first peptide] is an α-synuclein peptide, then [the second peptide] is an Aβ peptide, [ Each of Linker 1], [Linker 2], and [Cys] are optional, and [Linker 1] and [Linker 2] are the same or different linkers.

In certain embodiments, the dual alpha-synuclein polypeptide is as follows: [Cys]-[ Linker 2]-[ First Peptide ]-[ Linker 1]-[ Second Peptide ] where, if [First Peptide] peptide] is an Aβ peptide, then [second peptide] is an α-synuclein peptide, and if [first peptide] is an α-synuclein peptide, then [second peptide] is an Aβ peptide, and [links to] Each of [Linker 1], [Linker 2], and [Cys] is optional, and [Linker 1] and [Linker 2] are the same or different linkers.

Examples of A[beta] peptides include any of SEQ ID NOs 3-38 or 121-176.

Examples of alpha-synuclein peptides include any of SEQ ID NOs: 39-109.

[ Linker 1 ] is optionally present, and when present, it may be a cleavable linker. The cleavable linker, if present, can be 1 to 10 amino acids in length. In some embodiments, the linker comprises about 1 to 10 amino acids, about 1 to 9 amino acids, about 1 to 8 amino acids, about 1 to 7 amino acids, about 1 to 6 amino acids amino acid, about 1 to 5 amino acids, about 1 to 4 amino acids, about 1 to 3 amino acids, about 2 amino acids, or one (1) amino acid. In some embodiments, the cleavable linker is 1 amino acid, 2 amino acids, 3 amino acids, 4 amino acids, 5 amino acids, 6 amino acids, 7 amines amino acids, 8 amino acids, 9 amino acids, or 10 amino acids. In some embodiments, the linker can be a cleavable linker having an amino acid sequence selected from the group consisting of: arginine-arginine (Arg-Arg), arginine-valine-sperm Amino acid-arginine (Arg-Val-Arg-Arg; SEQ ID NO: 113), valine-citrulline (Val-Cit), valine-arginine (Val-Arg), valine Acid-lysine (Val-Lys), valine-alanine (Val-Ala), phenylalanine-lysine (Phe-Lys), glycine-alanine-glycine-alanine (Gly -Ala-Gly-Ala; SEQ ID NO: 114), alanine-glycine-alanine-glycine (Ala-Gly-Ala-Gly; SEQ ID NO: 115), or lysine-glycine -Lysine-glycine (Lys-Gly-Lys-Gly; SEQ ID NO: 116).

[ Linker 2 ] is optionally present, and when present, is the linker that couples the polypeptide to the carrier. The linker, if present, can be 1 to 10 amino acids in length. In some embodiments, the linker comprises about 1 to 10 amino acids, about 1 to 9 amino acids, about 1 to 8 amino acids, about 1 to 7 amino acids, about 1 to 6 amino acids amino acid, about 1 to 5 amino acids, about 1 to 4 amino acids, about 1 to 3 amino acids, about 2 amino acids, or one (1) amino acid. In some embodiments, the linker is 1 amino acid, 2 amino acids, 3 amino acids, 4 amino acids, 5 amino acids, 6 amino acids, 7 amino acids , 8 amino acids, 9 amino acids, or 10 amino acids. In some embodiments, the amino acid composition of the linker can mimic the composition of linkers found in native multidomain proteins, wherein certain amino acids are found in natural linkers compared to their abundance in the intact protein Over-present, under-present, or equally present. For example, threonine (Thr), serine (Ser), proline (Pro), glycine (Gly), aspartic acid (Asp), lysine (Lys), glutamine Acid (Gln), aspartic acid (Asn), arginine (Arg), phenylalanine (Phe), glutamic acid (Glu), and alanine (Ala) are overrepresented in natural linkers. In contrast, isoleucine (Ile), tyrosine (Tyr), tryptophan (Trp), and cysteine (Cys) were deficient. Typically, overrepresented amino acids are polar uncharged or charged residues that make up about 50% of the naturally encoded amino acids, and Pro, Thr, and GIn are the best amino acids for natural linkers. In some embodiments, the amino acid composition of the linker can mimic the composition of linkers typically found in recombinant proteins, which can generally be classified as flexible or rigid linkers. For example, flexible linkers found in recombinant proteins are typically composed of small, non-polar (eg, Gly) or polar (eg, Ser or Thr) amino acids, the small size of which provides flexibility and allows for the attachment of functional domains the movement. Incorporation of Ser or Thr, for example, can maintain the stability of the linker in aqueous solution by forming hydrogen bonds with water molecules, and thus can reduce the interaction between the linker and the immunogen. In some embodiments, the linker comprises an extension of Gly and Ser residues ("GS" linker). Examples of widely used flexible linkers are (Gly-Gly-Ser)n, (Gly-Gly-Gly-Ser)n (SEQ ID NO: 177) or (Gly-Gly-Gly-Gly-Ser)n (SEQ ID NO: 178), wherein n=1-3. Adjusting the number of copies "n" can optimize the linker to achieve sufficient separation of functional immunogenic domains, eg, to maximize immunogenic response. Many other flexible linkers have been designed for use with recombinant fusion proteins that can be used herein. In some embodiments, the linker may be rich in small or polar amino acids such as Gly and Ser, but also contain other amino acids such as Thr and Ala to maintain flexibility, and polar amino acids such as Lys and Glu acid to improve solubility. See, eg, Chen, X. et al., "Fusion Protein Linkers: Property, Design and Functionality" Adv Drug Deliv Rev. , 15; 65(10): 1357-1369 (203). In certain embodiments, the linker, when present, can be an amino acid sequence selected from the group consisting of: GG, GGG, KK, KKK, AA, AAA, SS, SSS, GAGA (SEQ ID NO: 114 ), AGAG (SEQ ID NO: 115) and KGKG (SEQ ID NO: 116).

[ Cys ] is optional and may assist in binding the polypeptide to the carrier. When present, Cys can be in the C-terminal portion of the polypeptide, or in the N-terminal portion of the polypeptide.

Examples of [first peptide]-[linker 1]-[second peptide]-[linker 2]-[Cys] dual Aβ/α-synuclein polypeptides of the present invention include the following: Table 1 Immunogen Assay ID A beta sequence Aβ SEQ ID NO. endopeptidase linker Alpha - synuclein sequence Alpha - synuclein SEQ ID NO. C -terminal connector Cys Immunogen SEQ ID NO 11 DAEFRHD 06 RR PDNEAYE 55 GG C 110 12 DAEFRHD 06 RR DPDNEAYE 48 GG C 111

Polypeptide Immunogen

Aβ peptides, α-synuclein peptides and dual Aβ/α-synuclein polypeptides are immunogens according to the present invention. In some embodiments, the peptides and dual A[beta]/[alpha]-synuclein polypeptides can be linked to a suitable carrier to help elicit an immune response. Thus, one or more of the peptides and dual A[beta]/[alpha]-synuclein polypeptides of the invention can be linked to a carrier. For example, each of the Aβ peptide, α-synuclein peptide, and Aβ/α-synuclein polypeptide can be linked to a carrier (eg, Gly- Gly, Ala-Ala, Lys-Lys, Ser-Ser, Gly-Ala-Gly-Ala (SEQ ID NO: 114), Ala-Gly-Ala-Gly (SEQ ID NO: 115) or Lys-Gly-Lys- Gly (SEQ ID NO: 116)). In certain embodiments, dual A[beta]-[alpha]-synuclein polypeptides can be linked to a suitable carrier using a C-terminal cysteine, to allow interposition between the peptide and the carrier or the dual A[beta]/[alpha]-synuclein polypeptide and the carrier linkers are provided. In certain embodiments, dual A[beta]-[alpha]-synuclein polypeptides can be linked to a suitable carrier using an N-terminal cysteine to provide a linker between the peptide and the carrier.

Suitable carriers include, but are not limited to, serum albumin, keyhole hemocyanin, immunoglobulin molecules, thyroglobulin, ovalbumin, tetanus toxoid, or toxins from other pathogenic bacteria, such as diphtheria toxoid ( For example CRM197), Escherichia coli ( E. coli ) , cholera or Helicobacter pylori ( H. pylori ), or sterile toxin derivatives . T cell epitopes are also suitable carrier molecules. Some conjugates can be made by attaching the peptide immunogens of the invention to immunostimulatory polymer molecules such as tripalmitoyl-S-glycerol cysteine (Pam3Cys), mannan (mannose polymer) or glucosamine Glycans (β1-2 polymers)), cytokines (eg IL-1, IL-1 alpha and beta peptides, IL-2, γ-INF, IL-10, GM-CSF) and chemotactic mediators elements such as MIP1-α and MIP1-β and RANTES. Other vectors include virus-like particles. In some compositions, the immunogenic peptide can also be linked to the carrier by chemical cross-linking. Techniques for attaching the immunogen to the carrier include the use of 3-(2-pyridylthio)propionic acid N-butadiimide (SPDP) and 4-(N-maleimidomethyl) disulfide bond (if the peptide does not have sulfhydryl groups, this can be provided by the addition of cysteine residues). These reagents generate disulfide bonds between themselves and peptide cysteine residues on a protein, and amides via the epsilon-amino group on lysine or other free amine groups in other amino acids key. In some embodiments, chemical cross-linking can include the use of SBAP (butanediimide 3-(bromoacetamido)propionate), which is used for via N-hydroxybutanediimide (NHS) ester A short (6.2 Angstrom) crosslinker for amine-sulfhydryl bonding with a bromoacetyl reactive group. A variety of such disulfide/amide formers are described in Jansen et al., "Immunotoxins: Hybrid Molecules Combining High Specificity and Potent Cytotoxicity" Immunological Reviews 62:185-216 (February 1982). Other bifunctional couplers form thioethers rather than disulfide bonds. Many of these thioether formers are commercially available and include 6-maleimidohexanoic acid, 2-bromoacetic acid and 2-iodoacetic acid, 4-(N-maleimide (methyl) cyclohexane-1-carboxylic acid reactive ester. The carboxyl group can be activated by combining it with succinimide or 1-hydroxy-2-nitro-4-sulfonic acid, sodium salt. Virus-like particles (VLPs), also known as pseudovirions or virus-derived particles, represent a plurality of viral capsids and/or envelope proteins capable of self-assembly in vivo into VLPs with defined spherical symmetry The subunit structure of replicas (Powilleit et al., (2007) PLoS ONE 2(5):e415). Alternatively, the peptide immunogen can be linked to at least one artificial T-cell epitope capable of binding to a majority of MHC class II molecules, such as a pan-DR epitope ("PADRE"). Pan DR binding peptides (PADRE) are described in US 5,736,142, WO 95/07707 and Alexander et al, Immunity , 1:751-761 (1994).

Active immunogens can be presented in multimeric form, wherein multiple copies of the immunogen (peptides of polypeptides) are presented on a carrier as a single covalent molecule. In some embodiments, the carrier includes various forms of dual A[beta]/[alpha]-synuclein polypeptides. For example, an immunogenic dual Aβ/α-synuclein polypeptide may include polypeptides of Aβ antigen and α-synuclein in different orders, or may be in the presence or absence of intrapeptide linkers and/or linkages In the case of a linker to a vector.

In some compositions, the immunogenic peptide can also be represented as a fusion protein with a carrier. In certain compositions, the immunogenic peptide can be attached to the carrier at the amino terminus, the carboxy terminus, or internally. In some compositions, the carrier is CRM197. In some compositions, the carrier is diphtheria toxoid.

nucleic acid

The present invention also provides nucleic acids encoding any of the amyloid-β (Aβ) peptides and α-synuclein peptides as disclosed herein. Nucleic acid immunotherapy compositions as disclosed herein comprise, consist of, or consist essentially of: a first nucleic acid sequence encoding an amyloid-beta (Aβ) peptide as disclosed herein and encoding an alpha- The second nucleic acid sequence of the nuclein peptide. For example, the Aβ peptide is a sequence of 3 to 10 amino acid residues in length and derived from the first ten N-terminal residues of SEQ ID NO: 01, and the α-synuclein peptide is 3 to 8 in length amino acid and from the sequence of residues 81 to 140 of SEQ ID NO:02. Accordingly, nucleic acids encoding any of SEQ ID NOs: 3-38 can be combined with nucleic acids encoding any of SEQ ID NOs: 39-109 to provide immunogens and components of pharmaceutical compositions of the invention . Likewise, one or more nucleic acids encoding any of the A[beta] and [alpha]-synuclein sequences may include codons for the RR-N-terminal or -RR C-terminal dipeptide. In certain embodiments, the Aβ and α-synuclein peptide sequences can be encoded by the same nucleic acid sequence or by different nucleic acid sequences. In some embodiments, the nucleic acid sequence may also encode a linker and/or a C-terminal cysteine for attachment to a vector, as described herein. Additionally, when a single nucleic acid sequence encodes two peptides, the sequence may also encode an intrapeptide linker as described herein. The nucleic acid compositions (pharmaceutical compositions) described herein can be used in methods of treating or effecting the prevention and/or prevention of Alzheimer's disease. In another embodiment, the nucleic acid immunotherapy compositions as disclosed herein provide compositions for reducing pathogenic forms of Aβ and α-synuclein in an individual and/or in the individual's tissues. In some embodiments, Aβ and/or α-synuclein reduced by the immunotherapy composition is a pathological form of: extracellular plaque deposits of Aβ (eg, β-amyloid (Aβ); neuritis amyloid plaques) and/or alpha-synuclein (eg, flame neurofibrillary tangles of alpha-synuclein oligomers). In other embodiments, pathological indicators of neurodegenerative diseases and/or synuclein disorders are reduced by nucleic acid immunotherapy compositions. In another embodiment, the nucleic acid immunotherapy compositions as disclosed herein provide compositions for reducing brain A[beta] and brain alpha-synuclein.

Nucleic acids such as DNA encoding an immunogen and used as a vaccine may be referred to as a "DNA immunogen" or "DNA vaccine" because the encoded polypeptide is expressed in vivo following administration of the DNA. DNA vaccines are intended to induce antibodies in an individual against the protein of interest that it encodes by integrating the DNA encoding the protein of interest into a vector (plastid or virus); administering the vector to the individual; and The protein of interest is expressed in the subject to which the vector is administered to stimulate the subject's immune system. DNA vaccines remain in the body of the individual long after administration and continue to produce the encoded protein slowly. Therefore, excessive immune responses can be avoided. DNA vaccines can also be modified using genetic engineering techniques. Optionally, such nucleic acids further encode a signal peptide and can be expressed together with a signal peptide linked to the peptide. The coding sequence of the nucleic acid can be operably linked to regulatory sequences to ensure the performance of the coding sequence, such as promoters, enhancers, ribosome binding sites, transcription termination signals, and the like. Nucleic acids encoding A[beta] and [alpha]-synuclein can exist in isolated form or can be cloned into one or more vectors. Nucleic acids can be synthesized by, for example, solid state synthesis or PCR of overlapping oligonucleotides. Nucleic acids encoding Aβ and α-synuclein peptides and polypeptides can be joined into one phase with and without a linker or cleavable linker, and with or without a protein-based carrier Adjacent nucleic acids, eg, within an expression vector.

DNA is more stable than RNA, but involves some potential safety risks such as inducing anti-DNA antibodies, so in some embodiments, the nucleic acid can be RNA. RNA nucleic acids encoding immunogens and used as vaccines may be referred to as "RNA immunogens" or "RNA vaccines" or "mRNA vaccines" because the encoded polypeptide is expressed in vivo following administration of the RNA. A ribonucleic acid (RNA) vaccine can safely direct the cellular machinery of an individual to produce one or more polypeptides of interest. In some embodiments, the RNA vaccine may be non-replicating mRNA (messenger RNA) or virus-derived self-amplifying RNA. mRNA-based vaccines encode antigens of interest and contain 5' untranslated regions and 3' untranslated regions (UTRs), while self-amplifying RNAs encode not only antigens, but also viruses that enable intracellular RNA amplification and expression of large amounts of proteins replication mechanism. In vitro transcribed mRNA can be generated from linear DNA templates using T7, T3 or Sp6 phage RNA polymerases. The resulting product may contain an open reading frame encoding a peptide of interest as disclosed herein, flanking 5'-UTR and 3'-UTR sequences, a 5' cap, and a poly(A) tail. In some embodiments, the RNA vaccine can comprise trans-amplified RNA (see, eg, Beissert et al., Molecular Therapy 2020 Jan 28(1):119-128). In certain embodiments, RNA vaccines encode Aβ peptides and α-synuclein peptides as disclosed herein, and are capable of expressing Aβ and α-synuclein peptides, particularly in transfer to antigen presentation such as immature in the case of cells within cells. RNA may also contain sequences encoding other polypeptide sequences such as immunostimulatory elements. In some embodiments, the RNA of the RNA vaccine can be modified RNA. In the context of RNA, the term "modified" can include any modification of RNA that is not naturally present in RNA. For example, a modified RNA can refer to an RNA with a 5'cap; however, the RNA can contain other modifications. The 5' cap can be modified to have the ability to stabilize it when attached to RNA. In certain embodiments, another modification may be an extension or truncation of the naturally occurring poly(A) tail or a change in the 5' untranslated region (UTR) or 3' untranslated region. In some embodiments, RNA vaccines or, eg, mRNA vaccines, are formulated in an effective amount to generate an antigen-specific immune response in an individual. For example, an RNA vaccine formulation is administered to an individual to stimulate the individual's humoral and/or cellular immune system against Aβ and α-synuclein antigens, and thus may further comprise one or more adjuvants, diluents, carriers, and and/or excipients, and administered to an individual by any suitable route to elicit a protective and/or therapeutic immune response against Aβ and α-synuclein antigens.

Fundamental references for uncovering general methods of molecular biology include: Sambrook, J et al., Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Press, Cold Spring Harbor, N.Y., 1989; Ausubel, FM et al., Current Protocols in Molecular Biology, Vol. 2, Wiley-Interscience, New York, (current edition); Kriegler, Gene Transfer and Expression: A Laboratory Manual (1990); Glover, DM, eds., DNA Cloning: A Practical Approach, Vol. Volume II, IRL Press, 1985; Albers, B. et al., Molecular Biology of the Cell, 2nd ed., Garland Publishing, Inc., New York, N.Y. (1989); Watson, J D. et al., Recombinant DNA, pp. 2nd edition, Scientific American Books, New York, 1992; and Old, RW et al., Principles of Gene Manipulation: An Introduction to Genetic Engineering, 2nd edition, University of California Press, Berkeley, Calif. (1981).

Techniques for manipulating nucleic acids, such as creating mutations in sequences, sub-selection, labeling probes, sequencing, hybridization, and the like, are well described in the scientific and patent literature. See, e.g., Sambrook, ed., MOLECULAR CLONING: A LABORATORY MANUAL (2nd Edition), Vols. 1-3, Cold Spring Harbor Laboratory, (1989); CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, ed. Ausubel, John Wiley & Sons, Inc. , New York (1997); LABORATORY TECHNIQUES IN BIOCHEMISTRY AND MOLECULAR BIOLOGY: HYBRIDIZATION WITH NUCLEIC ACID PROBES, Part I, edited by Tijssen, Elsevier, N.Y. (1993).

Nucleic acids, vectors, capsids, polypeptides, and analogs thereof can be analyzed and quantified by any of a variety of general methods well known to those skilled in the art. Such methods include, for example, analytical biochemical methods such as NMR, spectrophotometry, radiography, electrophoresis, capillary electrophoresis, high performance liquid chromatography (HPLC), thin layer chromatography (TLC) and superdiffusion Chromatography; various immunoassays such as liquid or gel precipitin reaction, immunodiffusion, immunoelectrophoresis, radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), immunofluorescence assay, southern Southern analysis, Northern analysis, dot blot analysis, gel electrophoresis (e.g. SDS-PAGE), RT-PCR, quantitative PCR; other nucleic acid or target or signal amplification methods, radioactivity Labeling, scintillation counting and affinity chromatography.

pharmaceutical composition

Each of the peptides and immunogens described herein can be presented in the form of pharmaceutical compositions administered with pharmaceutically acceptable adjuvants and pharmaceutically acceptable excipients. The adjuvant increases the titer of the induced antibody and/or the binding affinity of the induced antibody compared to the peptide alone. A variety of adjuvants can be used in combination with the immunogens of the invention to elicit an immune response. Some adjuvants enhance the innate response to the immunogen without causing conformational changes to the immunogen that affect the qualitative form of the response. Adjuvants can be natural compounds, modified versions or derivatives of natural compounds, or synthetic compounds.

Some adjuvants include aluminium salts (such as aluminium hydroxide and aluminium phosphate), 3-de-O-phosphorylated monophosphoryl lipid A (MPL ^™ ) (see GB 2220211 (RIBI ImmunoChem Research Inc., Hamilton, Montana, presently). of Corixa). As used herein, MPL refers to both natural and synthetic versions of MPL. Examples of synthetic versions include ^PHAD® , 3D- ^PHAD® , and 3D(6A) ^-PHAD® (Avanti Polar Lipids, Alabaster, Alabama).

QS-21 is a triterpene glycoside or saponin isolated from the bark of the Quillaja saponaria Molina tree found in South America (see Kensil et al., Vaccine Design: The Subunit and Adjuvant Approach (Powell and Newman). ed., Plenum Press, NY, 1995)). QS-21 products include Stimulon® (Antigenics, Inc., New York, NY; now Agenus, Inc. Lexington, MA) and QS-21 vaccine adjuvant (Desert King, San Diego, CA). QS-21 has been disclosed, characterized and evaluated in US 5,057,540 and US 8,034,348, the disclosures of which are incorporated herein by reference. Additionally, QS-21 has been evaluated in various clinical trials at various doses. See NCT00960531 (clinicaltrials.gov/ct2/show/study/NCT00960531), Hüll et al., Curr Alzheimer Res . 2017 Jul;14(7):696-708 (Evaluation of 50 mcg QS-21 with different doses of vaccine ACC -001); Gilman et al, "Clinical effects of Abeta immunization (AN1792) in patients with AD in an interrupted trial" Neurology . 2005 May 10;64(9):1553-62; Wald et al, "Safety and immunogenicity of long HSV-2 peptides complexed with rhHsc70 in HSV-2 seropositive persons Vaccine” 2011;29(47):8520-8529; and Cunningham et al., “Efficacy of the Herpes Zoster Subunit Vaccine in Adults 70 Years of Age or Older”. NEJM . 2016 Sep 15;375(11):1019-32. QS-21 is used in FDA-approved vaccines, including SHINGRIX. SHINGRIX contains 50 mcg of QS-21. In certain embodiments, the amount of QS-21 is from about 10 μg to about 500 μg.

TQL1055 is an analog of QS-21 (Adjuvance Technologies, Lincoln, NE). Compared to QS-21, semi-synthetic TQL1055 is characterized by high purity, improved stability, reduced local tolerance, and reduced systemic tolerance. TQL1055 has been disclosed, characterized and evaluated in US20180327436 A1, WO2018191598 A1, WO2018200656 A1 and WO2019079160 A1, the disclosures of which are incorporated herein by reference. US20180327436 A1 teaches that 2.5 times more TQ1055 is superior compared to 20 μg QS-21, but more than 50 μg TQ1055 shows no improvement. However, unlike QS-21, there was no increase in the degree of weight loss or hemolysis of RBCs with increasing doses of TQL1055. WO2018200656 A1 teaches that at the optimal amount of TQ1055, the antigenic amount can be reduced and a good titer can be achieved. In certain embodiments, the amount of TQL1055 is from about 10 μg to about 500 μg.

Other adjuvants are oil-in-water emulsions such as squalene or peanut oil, optionally in combination with immunostimulants such as monophosphoryl lipid A (see Stoute et al , N. Engl . J. Med 336, 86-91 (1997)), pluronic polymers and killed mycobacteria. Ribi adjuvant is an oil-in-water emulsion. Ribi contains a metabolizable oil (squalene) emulsified with physiological saline containing Tween 80. Ribi also contains an improved mycobacterial product that acts as an immunostimulant and bacterial monophosphoryl lipid A. Other adjuvants may be CpG oligonucleotides (see WO 98/40100), interleukins (eg IL-1, IL-1 alpha and beta peptides, IL-2, gamma-INF, IL-10, GM- CSF), chemokines (eg MIP1-alpha and MIP1-beta and RANTES), saponins, RNA and/or TLR agonists (eg TLR4 agonists such as MPL and synthetic MPL molecules), aminoalkyl Aminoglucoside phosphates and other TLR agonists. The adjuvant may be administered with the active agent as a component of the therapeutic composition or may be administered separately before, concurrently with, or after administration of the therapeutic agent.

In various embodiments of the invention, the adjuvant is QS-21 (Stimulon™). In some compositions, the adjuvant is MPL. In certain embodiments, the amount of MPL is from about 10 μg to about 500 μg. In some compositions, the adjuvant is TQL1055. In certain embodiments, the amount of TQL1055 is from about 10 μg to about 500 μg. In some compositions, the adjuvant is QS21. In certain embodiments, the amount of QS21 is from about 10 μg to about 500 μg. In some compositions, the adjuvant is a combination of MPL and QS-21. In some compositions, the adjuvant is a combination of MPL and TQL1055. In some compositions, the adjuvant can be in a liposomal formulation.

Additionally, some embodiments of the invention may include a multiplex antigen presentation system (MAP). Multiple antigen-presenting peptide vaccine systems have been developed to avoid the adverse effects associated with conventional vaccines (ie, live, killed, or inactivated pathogens), carrier proteins, and cytotoxic adjuvants. Two main approaches have been used to develop multiple antigen-presenting peptide vaccine systems: (1) adding functional components such as T-cell epitopes, cell-penetrating peptides, and lipophilic moieties; and (2) using nanoparticles of defined size Materials such as self-assembling peptides, non-peptide dendrimers, and gold nanoparticles serve as synthetic methods for antigen presentation platforms. The sometimes poor immunogenicity of subunit peptide vaccines can be improved using the multiple antigen peptide (MAP) system. In the MAP system, multiple copies of the antigenic peptide are simultaneously bound to the α- and ε-amine groups of the non-immunogenic Lys-based dendritic architecture, helping to impart stability against degradation, thereby enhancing immune cell interaction. Molecular recognition and induction of stronger immune responses than small antigenic peptides alone. In some compositions, MAPs comprise Lys-based dendritic structures, helper T cell epitopes, immunostimulatory lipophilic moieties, cell penetrating peptides, free radical-induced polymerization, self-assembling nanoparticles (as antigens) presentation platform) and one or more of gold nanoparticles.

Pharmaceutical compositions for parenteral administration are preferably sterile and substantially isotonic, and manufactured under GMP conditions. Pharmaceutical compositions may be presented in unit dosage form (ie, for single administration of a dose). Pharmaceutical compositions can be formulated using one or more physiologically acceptable carriers, diluents, excipients or adjuvants. Formulations depend on the route of administration chosen. For injection, the peptides of the present invention may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hank's solution, Ringer's solution or physiological saline or acetic acid Salt buffer (to reduce discomfort at the injection site). The solutions may contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Alternatively, the peptide composition can be in lyophilized form for reconstitution with a suitable vehicle, eg, sterile pyrogen-free water, before use.

The peptide (and optionally a vector fused to the peptide) can also be administered in the form of a nucleic acid encoding the peptide and expressed in situ in an individual. Nucleic acid segments encoding immunogens are typically linked to regulatory elements, such as promoters and enhancers, that enable expression of the DNA segment in the intended target cells in an individual. For expression in blood cells, promoters and enhancer elements from eg light or heavy chain immunoglobulin genes or CMV major intermediate early promoters and enhancers are suitable for directing expression as required to induce an immune response. The linked regulatory elements and coding sequences are typically cloned into the vector.

DNA and RNA can be delivered in naked form (ie, in the absence of colloidal or encapsulating materials). Alternatively, various viral vector systems can be used, including retroviral systems (see, eg, Boris-Lawrie and Teumin, Cur . Opin . Genet . Develop . 3(1): 102-109 (1993)); adenoviral vectors (see, eg, Bett et al . , J. Virol . 67(10); 5911-21 (1993)); adeno-associated viral vectors (see, eg, Zhou et al . , J. Exp . Med . 179(6):1867-75 (1994)) ; viral vectors from the Pox family, including vaccinia virus and fowlpox virus; viral vectors from alphaviruses, such as those from Sindbis and Semliki Forest Virus ( See, eg, Dubensky et al, J. Virol . 70(1):508-519 (1996)); Venezuelan equine encephalitis virus (see US 5,643,576) and baculoviruses such as vesicular stomatitis virus (see WO 96/34625) and papilloma virus (WO 94/12629; Ohe et al, Human Gene Therapy 6(3):325-333 (1995); and Xiao and Brandsma, Nucleic Acids . Res . 24 ( 13): 2620-2622 (1996)).

DNA and RNA encoding immunogens or vectors containing them can be encapsulated in liposomes, nanoparticles or lipoprotein complexes. Suitable other polymers include, for example, protamine liposomes, polysaccharide particles, cationic nanoemulsions, cationic polymers, cationic polymer liposomes, cationic lipid nanoparticles, cationic lipids, cholesterol nanoparticles, cationic lipid-cholesterol, PEG nanoparticles or dendritic polymer nanoparticles. Other suitable lipids and related analogs are described by US 5,208,036, US 5,264,618, US 5,279,833 and US 5,283,185, each of which is incorporated herein by reference in its entirety. Carriers and DNA encoding immunogens can also be adsorbed to or associated with particulate carriers, examples of which include polymethylmethacrylate polymers and polylactic acid and poly(lactide-co-glycolide) (see For example McGee et al, J. Micro Encap . 1997 Mar-Apr; 14(2):197-210).

The pharmaceutically acceptable carrier composition may also include additives including, but not limited to, water, pharmaceutically acceptable organic solvents, collagen, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymers , Sodium carboxymethyl cellulose, sodium polyacrylate, sodium alginate, water-soluble polydextrose, sodium carboxymethyl starch, pectin, methyl cellulose, ethyl cellulose, xanthan gum, gum arabic (gum arabic), casein, agar, polyethylene glycol, diglycerol, glycerol, propylene glycol, paraffin fat, paraffin stearyl alcohol, stearic acid, human serum albumin, mannitol, sorbitol, lactose and can Surfactants accepted as pharmaceutical additives.

Individuals eligible for treatment

The presence of Aβ plaques and/or neurofibrillary tangles has been found in several diseases, including Alzheimer's disease, Down's syndrome, mild cognitive impairment, amyloid cerebrovascular disease, post-encephalitic Barking postencephalitic parkinsonism, post-traumatic dementia or dementia pugilistica, Pick's disease, Niemann-Pick disease type C, supranuclear palsy, Frontotemporal dementia, frontotemporal degeneration, argentophilic granular dementia, amyotrophic lateral sclerosis/parkinsonism dementia complex of Guam, corticobasal degeneration (CBD), Lewy Body Dementia, Lewy Body Variation of Alzheimer's Disease (LBVAD), Chronic Traumatic Encephalopathy (CTE), Parkinson's Disease, Progressive Supranuclear Palsy (PSP), Dry age-related macular degeneration (AMD) and inclusion body myositis.

The compositions and methods of the present invention can be used to treat or prevent any of these diseases. Due to the broad correlation between neurological disease and A[beta] and/or alpha-synuclein, the compositions and methods of the present invention can be used to treat or prevent neurological disease compared to the mean of individuals without neurological disease. Any subject showing elevated levels of A[beta] and/or [alpha]-synuclein (eg, in CSF). The compositions and methods of the present invention may also be used to treat or prevent neurological disease in individuals with neurological disease-associated mutations in A[beta] and/or [alpha]-synuclein. The methods are particularly useful for treating or preventing Alzheimer's disease.

Individuals who can receive treatment include individuals at risk of developing the disease but not showing symptoms, as well as patients currently showing symptoms, including untreated individuals who have not previously received treatment for the disease. Individuals at risk of developing the disease include individuals in the elderly population, asymptomatic individuals with Aβ and/or α-synuclein disorders, and individuals at known risk for genetic disease. Such individuals include individuals with relatives who have experienced the disease, as well as individuals at risk determined by analysis of genetic or biochemical markers. Genetic markers of risk include mutations in A[beta] and/or [alpha]-synuclein, as well as mutations in other genes associated with neurological diseases. For example, heterotypic combinations and even more ApoE4 dual genes in homotypic combinations are associated with risk of Alzheimer's disease (AD). Other markers of Alzheimer's risk include mutations in the APP gene, particularly at positions 717 and 670 and 671 known as Hardy and Swedish mutations, respectively, Mutations in presenilin genes, PS1 and PS2, family history of AD, hypercholesterolemia or atherosclerosis. Individuals currently suffering from Alzheimer's disease can be identified by PET imaging, by characteristic dementia and the presence of the risk factors described above. Additionally, there are many diagnostic tests that can be used to identify individuals with AD. These tests include measurements of CSF or blood alpha-synuclein and Abeta42 levels. Elevated alpha-synuclein and decreased A[beta]42 levels indicate the presence of AD. Some mutations are associated with Parkinson's disease, such as Ala30Pro or Ala53Thr, or mutations in other genes are associated with Parkinson's disease, such as leucine-rich repeat kinases (LRRK2 or PARK8). Whether an individual suffers from any of the neurological diseases mentioned above can also be diagnosed by the criteria of DSM IV TR.

In asymptomatic individuals, treatment can be initiated at any age (eg, 10 years, 20 years, 30 years or older). Typically, however, treatment does not need to be started until the individual reaches 20, 30, 40, 50, 60, 70, 80 or 90 years of age. Treatment usually requires multiple doses over a period of time. Treatment can be monitored by analyzing antibody levels over time. If the response declines, administration of a booster dose is indicated. For patients with underlying Down syndrome, treatment can be initiated prenatally or shortly after birth by administering the therapeutic agent to the mother.

Treatment methods and uses

The present invention provides methods of inhibiting or reducing the aggregation of A[beta] or [alpha]-synuclein in individuals suffering from or at risk of developing a neurodegenerative disease such as Alzheimer's disease. The methods include administering to an individual a composition as disclosed herein. A therapeutically effective amount is one that, when provided within an effective period, will achieve the desired immunological or clinical effect. Dosage regimens can be adjusted to provide optimal therapeutic response. For example, several divided doses may be administered at set intervals (eg, one week, one month), or the dose may be proportionally reduced as the therapeutic situation warrants.

In prophylactic applications, a regimen (dosage, frequency, and route of administration) that is effective in reducing the risk, lessening the severity, or delaying the onset of at least one sign or symptom of a disease is directed toward a predisposing disease (eg, Alzheimer's disease). ) or an individual at risk of developing a disease is administered a composition described herein. In particular, the regimen is effective in inhibiting or delaying the formation of Aβ plaques, and/or inhibiting or delaying alpha-synuclein disorders, and/or inhibiting or delaying its toxic effects, and/or inhibiting or delaying its toxic effects in the brain Development of behavioral deficits. In therapeutic applications, a subject suspected of having a disease (eg, Alzheimer's disease) is administered a regimen (dosage, frequency and route of administration) effective to ameliorate at least one sign or symptom of the disease or at least inhibit its further progression Or patients with pre-existing disease are administered the compositions described herein. In particular, the regimen is preferably effective in reducing the level of, or at least inhibiting further increases in, toxicity and/or behavioral deficits associated with A[beta] plaques and/or synuclein disorders.

If treated individuals achieve results that are more favorable than the average outcome in a control population of comparable individuals not treated by the methods of the invention, or if in controlled clinical trials (eg, Phase II, Phase II/III or A regimen can be considered therapeutically or prophylactically effective in a phase III trial) at the p<0.05 or 0.01 or even 0.001 level in treated subjects showing more favorable results than control subjects.

Effective doses vary depending on many different factors, such as the mode of administration, the target site, the physiological state of the patient, whether the patient is an ApoE carrier, whether the patient is a human or animal, other drugs administered, and whether the treatment is prophylactic or therapeutic sex.

In some embodiments, the effective amount is a total dose of 25 μg to 1000 μg or 50 μg to 1000 μg. In some embodiments, the effective amount is a total dose of 100 μg. In some embodiments, the effective amount is a total of two doses of 25 μg administered to the subject. In some embodiments, an effective amount is a total of two doses of 100 μg administered to an individual. In some embodiments, an effective amount is a total of two doses of 400 μg administered to an individual. In some embodiments, an effective amount is a total of two doses of 500 μg administered to an individual. In some embodiments, RNA (eg, mRNA) vaccines are administered to an individual by intradermal, intramuscular injection, or by intranasal administration.

In some embodiments, the amount of the agent for active immunotherapy ranges from 1 to 1,000 micrograms (μg), or 0.1 to 500 μg, or 10 to 500 μg, or 50 to 250 μg per patient and for humans Administration can be 1 to 100 or 1 to 10 μg per injection. The timing of injections can vary significantly from once a day to once a week, once a month, once a year, once a decade. A typical regimen consists of immunization followed by booster injections at time intervals, such as 6-week intervals or two months. Another regimen consists of immunization followed by one or more booster injections 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months later. Another regimen requires injections every two months for life. Alternatively, occasional booster injections may be administered as indicated by monitoring of the immune response. The frequency of administration may be one or more times, as long as the side effects are within a clinically acceptable range.

In some embodiments, a composition or method as disclosed herein comprises administering to an individual a nucleic acid vaccine comprising one or more DNA or RNA polymers having open reading frames encoding a first peptide and a second peptide Nucleotides, wherein a nucleic acid vaccine is administered to an individual at a dose between 10 μg/kg and 400 μg/kg. In some embodiments, the dose of RNA polynucleotide is 1 to 5 μg, 5 to 10 μg, 10 to 15 μg, 15 to 20 μg, 10 to 25 μg, 20 to 25 μg, 20 to 50 μg per dose , 30 to 50 μg, 40 to 50 μg, 40 to 60 μg, 60 to 80 μg, 60 to 100 μg, 50 to 100 μg, 80 to 120 μg, 40 to 120 μg, 40 to 150 μg, 50 to 150 μg , 50 to 200 μg, 80 to 200 μg, 100 to 200 μg, 120 to 250 μg, 150 to 250 μg, 180 to 280 μg, 200 to 300 μg, 50 to 300 μg, 80 to 300 μg, 100 to 300 μg , 40 to 300 μg, 50 to 350 μg, 100 to 350 μg, 200 to 350 μg, 300 to 350 μg, 320 to 400 μg, 40 to 380 μg, 40 to 100 μg, 100 to 400 μg, 200 to 400 μg or 300 to 400 μg. In some embodiments, the nucleic acid is administered to the individual by intradermal or intramuscular injection. In some embodiments, the nucleic acid is administered to the individual on day zero. In some embodiments, the subject is administered a second dose of nucleic acid on day seven, or day fourteen, or day twenty-first.

The compositions described herein are preferably administered via a peripheral route (that is, the administered compositions elicit a stable immune response and/or induce a population of antibodies across the blood-brain barrier to reach the brain, spinal cord, or eyes route to the desired site in the . For peripheral disease, the induced antibodies leave the vasculature to reach the intended peripheral organs. Routes of administration include oral, subcutaneous, intranasal, intradermal or intramuscular. Some routes of active immunization are subcutaneous and intramuscular. Intramuscular and subcutaneous administration can be performed at a single site or at multiple sites. Intramuscular injections are most commonly given in the arm or leg muscle. In some methods, the agent is injected directly into the specific tissue in which the deposit has accumulated.

The amount of the dose administered can be adjusted to produce a more stable immune response (eg, a higher titer). For acute conditions or acute exacerbations of chronic conditions, 1 to 10 doses are usually sufficient. Sometimes a single bolus injection dose in divided form, as appropriate, is sufficient for an acute condition or acute exacerbation of a chronic condition. For chronic conditions, vaccines/immunotherapy as disclosed herein can be administered at regular intervals, eg, weekly, biweekly, monthly, quarterly, every six months, for at least 1, 5 or 10 years, or the patient's lifetime.

An effective amount of the DNA- or RNA-encoded immunogen may be between about 1 nanogram and about 1 gram per kilogram of recipient body weight, or between about 0.1 μg/kg and about 10 mg/kg, or about about Between 1 μg/kg and about 1 mg/kg. Dosage forms suitable for internal administration preferably contain (for the latter dosage range) from about 0.1 μg to 100 μg of active ingredient per unit. The active ingredient may range from 0.5 to 95% by weight, based on the total weight of the composition. Alternatively, an effective dose of antigen-loaded dendritic cells is between about ¹⁰⁴ and ¹⁰⁸ cells. Those skilled in immunotherapy should be able to adjust these doses without undue experimentation.

The nucleic acid composition can be administered in a convenient manner, such as injection by a convenient and effective route. Routes may include, but are not limited to, intradermal "gene gun" delivery or intramuscular injection. The modified dendritic cells are administered by subcutaneous, intravenous or intramuscular routes. Other possible routes include oral, intrathecal, inhalation, transdermal or rectal administration.

Depending on the route of administration, the composition can be coated in a material to protect the compound from the action of enzymes, acids, and other natural conditions that may inactivate the compound. Therefore, it may be necessary to coat the composition with a material or to co-administer the composition with a material to prevent the composition from deactivating. For example, enzyme inhibitors of nucleases or proteases (eg, trypsin inhibitor, diisopropyl fluorophosphate, and aprotinin) or enzyme inhibitors such as liposomes (including water-in-oil-in-water emulsions) and conventional lipids body (Strejan et al , J. Neuroimmunol 7(1):27-41, 1984) in a suitable carrier.

The immunotherapeutic compositions disclosed herein may also be used in combination with other treatments for diseases associated with accumulation of Aβ or α-synuclein, such as anti-Aβ antibodies, such as those that specifically bind to those disclosed herein Antibodies to any of the Aβ epitopes. For example, aducanumab or any of the antibodies disclosed in, eg, US Patent Publication No. 2010/0202968 and US Patent No. 8,906,367, and/or anti-alpha-synuclein antibodies such as Antibodies that specifically bind to any of the alpha-synuclein epitopes disclosed herein), ABBV-8E12, gosuranemab, zagotenemab, RG-6100 ; Nucleoprotein antibodies, such as antibodies that specifically bind to any of the alpha-synuclein epitopes disclosed herein, or antibodies that bind to compounds and/or other alpha-synuclein proteins, such as PRX002/ RO7046015, PRX002/RG7935 (Prasinezumab), NPT200-11/UCB0599, NPT088, BIIB054 (Cinpanemab), ABBV-0805, MEDI-1341, NPT088, Lu AF82422. In some combination therapy approaches, the patient receives passive immunotherapy prior to the active immunotherapy approaches disclosed herein. In other methods, patients receive passive and active immunotherapy during the same treatment period. Alternatively, patients can receive active immunotherapy before passive immunotherapy. Combinations may also include small molecule therapies and non-immunogenic therapies such as RAZADYNE ^® (galantamine), EXELON ^® (rivastigmine) and ARICEPT ^® (donepezil) and other compositions for improving the function of nerve cells in the brain.

The compositions of the present invention can be used in the manufacture of medicaments for use in the treatment regimens described herein.

treatment plan

The desired results of the methods of treatment as disclosed herein vary depending on the disease and patient profile, and can be determined by those skilled in the art. Desired outcomes include improvement in the patient's health status. Typically, desired outcomes include measurable indices such as reduction or clearance of pathological amyloid fibrils, reduction or inhibition of amyloid aggregation and/or amyloid fibril deposition, and reduction in pathological and/or aggregation The immune response to such amyloid fibrils is increased. Desired results also include amelioration of amyloid disease-specific symptoms. As used herein, relative terms such as "improved," "increased," or "reduced" indicate relative to a control (such as a measurement in the same individual prior to initiating a treatment described herein, or in a control measurements in individuals or groups). Control individuals were individuals who had the same amyloid-like disease as the treated individuals and were approximately the same age as the treated individuals (to ensure similar disease stages in the treated and control individuals), but who had not received the Treatment by the disclosed immunotherapy/vaccine formulations. Alternatively, control individuals are healthy individuals approximately the same age as the treated individuals. Changes or improvements in response to treatment are generally statistically significant and can be considered significant when described by a p-value of less than or equal to 0.1, less than 0.05, less than 0.01, less than 0.005, or less than 0.001.

Effective doses of compositions as disclosed herein for treating an individual vary depending on a number of different factors, including the mode of administration, the site of interest, the physiological state of the patient, whether the patient is human or animal, other drugs administered (if any). ) and whether treatment is prophylactic or curative. Therapeutic doses can be analyzed in a titrated fashion to optimize safety and efficacy. The amount of immunogen may also depend on whether adjuvant is also administered, with higher doses being required in the absence of adjuvant. The amount of immunogen used for administration sometimes ranges from 1 to 500 μg per patient and, for human administration, more usually ranges from 5 to 500 μg per injection. Occasionally, higher doses of 1 to 2 mg per dose are used. Typically, about 10, 20, 50 or 100 μg is used per human dose. The timing of dosing can vary significantly from once a day to once a year to once a decade. On any given day a dose of the immunogen is provided, if an adjuvant is also administered, the dose is greater than 1 microgram per patient and usually greater than 10 microgram per patient, and in the absence of adjuvant greater than 10 microgram per patient and usually greater than 100 micrograms/patient. A typical regimen consists of immunization followed by booster doses at 6-week intervals. Another regimen consists of immunization followed by administration of booster doses 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months later. Another regimen requires doses to be administered every two months for life. Alternatively, booster doses may be administered occasionally as indicated by immune response monitoring.

When administered in combination with a second treatment for Alzheimer's disease, such as Razadyne® (galantamine), Exelon® (rivastigmine), and Aricept® (donepezil), may vary depending on the product A second treatment is administered according to treatment with the composition of the present invention as needed.

set

The present invention also provides kits (eg, containers) comprising the compositions disclosed herein and related materials, such as instructions for use (eg, drug instructions). Instructions for use may contain, for example, instructions for administration of the composition and, optionally, one or more other agents. Containers for peptide and/or nucleic acid compositions can be unit doses, bulk packages (eg, multi-dose packages), or sub-unit doses.

Package insert means the instructions normally included in the package of a commercially available therapeutic product that contain information about the indications, usage, dosage, administration, contraindications and/or warnings related to the use of such therapeutic product. The kit may also include a second container comprising a pharmaceutically acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution, and dextrose solution. It may also include other materials required from a commercial and user standpoint, including other buffers, diluents, filters, needles and syringes.

use

Each of the peptides, polypeptides, immunogens, and pharmaceutical compositions described herein can be used to treat one or more diseases as described herein. Additionally, each of the peptides, polypeptides, immunogens, and pharmaceutical compositions described herein can be used in methods for treating one or more diseases as described herein. Each of the peptides, polypeptides, immunogens and pharmaceutical compositions described herein can be used in a method for the manufacture of a medicament for treating or for treating one or more diseases as described herein.

The following is provided for illustrative purposes only, and is not intended to limit the scope of the invention described above in the broad sense.

All US and international patent applications identified herein are incorporated by reference in their entirety. example

Example 1 : Animal immunization

On days 0, 14, 42 and 70, two sites of Swiss Webster mice were injected subcutaneously with 100 μl of the test immunogen (200 μl total). Test immunogens were prepared by combining 25 μg of test immunogen and 25 μg of QS21 adjuvant in 200 μl of Phosphate Buffered Saline (PBS). On days 21, 49 and 77, mice were bled by tail-cutting and 50 μl of blood was collected and subsequently processed into serum. The immunogens tested included DAEFRHDRRPDNEAYEGGC (SEQ ID NO: 110) and DAEFRHDRRDPDDNEAYEGGC (SEQ ID NO: 111). The immunogen contains an Aβ peptide, an α-synuclein peptide, a C-terminal linker, and a C-terminal cysteine (ie, -Gly-Gly-Cys-) and is linked via a C-terminal cysteine with maleic acid. CRM-197 coupling of diimide linkages.

On days 0, 21, 49 and 77, guinea pigs were injected intramuscularly with 50 μg of the test immunogen, 200 μl Addavax containing 25 μg QS21. Bloodletting was performed 7 days after immunization. The immunogens tested included DAEFRHDRRPDNEAYEGGC (SEQ ID NO: 110) and DAEFRHDRRDPDDNEAYEGGC (SEQ ID NO: 111). The immunogen contains an Aβ peptide, an α-synuclein peptide, a C-terminal linker, and a C-terminal cysteine (ie, -Gly-Gly-Cys-) and is linked via a C-terminal cysteine with maleic acid. CRM-197 coupling of diimide linkages.

At the start of the study, female guinea pigs were at least 5 weeks old and weighed approximately 350 to 500 g. Appropriate for animal husbandry and care in an accredited facility in accordance with U.S. Department of Agriculture's (USDA) and International Assessment and Accreditation of Laboratory Animal Care (AAALAC) guidelines Animal housing and research procedures.

The immunogen concentration was 0.5 mg/ml. Prior to each administration of the test immunogen, an approximately 3 ^cm2 area on each hind limb was shaved and rinsed with ethanol to visualize the injection site. Each animal received a dose of 200 microliters (0.25 micrograms/microliter) of the test immunogen divided into two separate sites with 100 microliters per injection per site (ie, animals received 100 microliters containing 50 micrograms of immunogen). μl PBS + 100 μl Addavax containing 25 μg QS-21). A 25G-27G needle was inserted intramuscularly into the hind limb to a depth of about 0.25 to 0.5 cm and injected at 100 microliters per site. For each administration, injection sites were alternated between four separate sites on each hind limb separated by at least 2 cm.

Example 2 : Measurement of antibody titer

Guinea pig whole blood samples of 250-350 μl each were collected via the jugular vein at weeks 1, 4, 8, and 12 and borrowed at weeks 1, 3, 7, and 11. 50 microliters of whole blood samples were collected each time by tail cutting the mice into clot activator tubes. The maximum volume of whole blood was collected via cardiac puncture into clot activator tubes at the end of the final collection week. All blood samples were allowed to clot for more than 30 minutes at room temperature, centrifuged at 3,000 RPM for 10 to 15 minutes at ambient temperature (approximately 20 to 25°C), and serum supernatants were transferred individually to clean frozen vials. Serum supernatants were stored frozen at -80°C (±12°C).

Aβ titer (mouse)

2 μg/ml of Aβ 1-28 monomer was spread on plates at 100 μl per well in PBS and incubated overnight at room temperature. Plates were blocked with 1% BSA in PBS for 1 hour. The plate was aspirated and 200 μl of PBS Tween with 0.1% BSA was added to column A. In row 1, negative mouse serum was added at 1/100, while the rest of the column contained 1/100 of the test serum. Serially dilute each column in the plate by ½ to obtain a dilution of 1/100 to 1/12800. Wells were incubated for 2 hours at room temperature, followed by washing, and a 1/5000 dilution of anti-mouse IgG HRP in PBS Tween with 0.1% BSA was prepared and 100 μl was then added to the washed wells. It was incubated for 1 hour and washed. OPD matrices were prepared using Thermo-Fisher OPD lozenges at 1 lozenge per 10 ml. Thermo-fisher substrate buffer was added at 1/10 and 100 μl was added to each well and incubated for 15 minutes. 50 μl of _{2N H2SO4} was added to stop the reaction, and the plate was read with a Molecular Devices Spectromax at 490 nM. The titer was defined as the dilution that provided 50% of the maximum OD and extrapolated if it was between dilutions.

Alpha-synuclein titer (mouse)

2 μg/ml of recombinant human alpha-synuclein was plated on plates at 100 μl per well in PBS and incubated overnight at room temperature. Plates were blocked with 1% BSA in PBS for 1 hour. The plate was aspirated and 200 μl of PBS Tween with 0.1% BSA was added to column A. In row 1, negative mouse serum was added at 1/100, while the rest of the column contained 1/100 of the test serum. Serially dilute each column in the plate by ½ to obtain a dilution of 1/100 to 1/12800. Wells were incubated for 2 hours at room temperature and then washed, and a 1/5000 dilution of anti-mouse IgG HRP in PBS Tween with 0.1% BSA was prepared and 100 μl was then added to the washed wells. It was incubated for 1 hour and washed. OPD matrices were prepared using Thermo-Fisher OPD lozenges at 1 lozenge per 10 ml. Thermo-fisher substrate buffer was added at 1/10 and 100 μl was added to each well and incubated for 15 minutes. 50 μl of _{2N H2SO4} was added to stop the reaction, and the plate was read with a Molecular Devices Spectromax at 490 nM. The titer was defined as the dilution that provided 50% of the maximum OD and extrapolated if it was between dilutions.

Aβ titer (guinea pig)

2 μg/ml of Aβ 1-28 monomer was spread on plates at 100 μl per well in PBS and incubated overnight at room temperature. Plates were blocked with 1% BSA in PBS for 1 hour. The plate was aspirated and 200 μl of PBS Tween with 0.1% BSA was added to column A. In row 1, negative guinea pig serum was added at 1/100, while the rest of the columns contained 1/100 of the test serum. Serially dilute each column in the plate by ½ to obtain a dilution of 1/100 to 1/12800. Wells were incubated for 2 hours at room temperature, followed by washing, and a 1/5000 dilution of anti-guinea pig IgG HRP in PBS Tween with 0.1% BSA was prepared and 100 μl was then added to the washed wells. It was incubated for 1 hour and washed. OPD matrices were prepared using Thermo-Fisher OPD lozenges at 1 lozenge per 10 ml. Thermo-fisher substrate buffer was added at 1/10 and 100 μl was added to each well and incubated for 15 minutes. 50 μl of _{2N H2SO4} was added to stop the reaction, and the plate was read with a Molecular Devices Spectromax at 490 nM. The titer was defined as the dilution that provided 50% of the maximum OD and extrapolated if it was between dilutions.

Alpha-synuclein titer (guinea pig)

2 μg/ml of recombinant human alpha-synuclein was plated on plates at 100 μl per well in PBS and incubated overnight at room temperature. Plates were blocked with 1% BSA in PBS for 1 hour. The plate was aspirated and 200 μl of PBS Tween with 0.1% BSA was added to column A. In row 1, negative guinea pig serum was added at 1/100, while the rest of the columns contained 1/100 of the test serum. Serially dilute each column in the plate by ½ to obtain a dilution of 1/100 to 1/12800. Wells were incubated for 2 hours at room temperature, followed by washing, and a 1/5000 dilution of anti-guinea pig IgG HRP in PBS Tween with 0.1% BSA was prepared and 100 μl was then added to the washed wells. It was incubated for 1 hour and washed. OPD matrices were prepared using Thermo-Fisher OPD lozenges at 1 lozenge per 10 ml. Thermo-fisher substrate buffer was added at 1/10 (100 μl to each well) and the mixture was incubated for 15 minutes. 50 μl of _{2N H2SO4} was added to stop the reaction, and the plate was read with a Molecular Devices Spectromax at 490 nM. The titer was defined as the dilution that provided 50% of the maximum OD and extrapolated if it was between dilutions.

Antibody titers observed in guinea pigs immunized as described above are shown in Table 1 . Immunization was performed with Addavax containing QS21. The reported titers are for blood after the second injection. These results are presented in Figure 1 .

surface 1 . through A beta and alpha - guinea pig immunized with synuclein immunogen ( GP ) Antibody titers in . immunogen GP1Aβ potency GP 2 Aβ potency GP 3 Aβ potency DAEFRHDRRPDNEAYEGGC (SEQ ID No: 110) 700 300 500 DAEFRHDRRDPDDNEAYEGGC (SEQ ID No: 111) 400 600 500 immunogen GP 1 α- synuclein titer GP 2 α- synuclein titer GP 3 α- synuclein titer DAEFRHDRRPDNEAYEGGC (SEQ ID No: 110) 100 300 300 DAEFRHDRRDPDDNEAYEGGC (SEQ ID No: 111) 400 300 350

Antibody titers observed in mice immunized as described above are shown in Table 2. Immunization with QS21. The reported titers are for blood after the third injection. These results are presented in FIG. 2 .

surface 2 . through A beta and alpha - Antibody titers in mice immunized with synuclein immunogen. immunogen mouse 1 Aβ potency mouse 2 Aβ potency mouse 3 Aβ potency mouse 4 Aβ potency DAEFRHDRRPDNEAYEGGC (SEQ ID No: 110) 3000 10000 - - DAEFRHDRRDPDDNEAYEGGC (SEQ ID No: 111) 2000 700 10000 2400 immunogen mouse 1 alpha- synuclein titer mouse 2α- synuclein titer mouse 3α- synuclein titer mouse 4 alpha- synuclein titer DAEFRHDRRPDNEAYEGGC (SEQ ID No: 110) 700 1000 - - DAEFRHDRRDPDDNEAYEGGC (SEQ ID No: 111) 12500 9000 6000 15000

Example 3 : Staining of Alzheimer's brain tissue with serum from animals immunized with vaccines as disclosed herein .

In the presence of sodium azide, tissue sections were placed in a solution of glucose oxidase and βD-glucose to block endogenous peroxidase. After tissue sections were prepared, the appropriate animal species secondary antibody and the DAKO DAB detection kit were used in two dilutions (1:300 and 1:1500) according to the manufacturer's instructions with the appropriate animal species secondary antibody as disclosed herein. The indicated sera from vaccinated animals were stained. Staining was performed using an automated Leica Bond Stainer. The results indicate whether serum from animals immunized with a vaccine as disclosed herein contains antibodies specific for Aβ and/or α-synuclein in human brain tissue of Alzheimer's patients .

in conclusion

Dual immunogenic Aβ-α-synuclein vaccine constructs were developed and shown to increase the balanced titers of Aβ and α-synuclein in mice, guinea pigs and cynomolgus monkeys. These antibodies are immunoreactive against both Aβ plaques and neurofibrillary α-synuclein in human AD brain slices, and block the binding of soluble Aβ aggregates (oligomers) to neurons without Triggers T cell responses against Aβ or α-synuclein. These results provide support for the development of a single-dose dual immunogenic vaccine capable of targeting both Aβ and α-synuclein in pathogenic forms. These results provide support for the development of dual Aβ-α-synuclein vaccines capable of targeting both pathogenic Aβ and α-synuclein to prevent and/or treat AD.

While various specific embodiments of the invention have been described herein, it is to be understood that the invention is not limited to these specific embodiments and that various changes may be effected by those skilled in the art without departing from the scope and spirit of the invention or modified.

SEQ ID NO:02 - α-突觸核蛋白同功異型物NACP140 [智人] NCBI參考序列：NP_000336.1

Aβ 免疫原：

α - 突觸核蛋白免疫原

DAEFRHDRRX ₁PDNEAYEXXC     (SEQ ID NO:112)，其中X ₁為視情況存在，且若存在，則X ₁為D，且其中XX及C獨立地為視情況存在，且若存在，則XX可為GG、AA、KK、SS、GAGA、AGAG或KGKG。

In various embodiments of the peptides described herein, the peptide may comprise, consist of, or consist essentially of the recited sequence. Accordingly, the following sequences incorporated into the present invention (see Table 3) can be part of a composition comprising amyloid-beta (Abeta)-like peptides and alpha-synuclein (poly)peptides as disclosed herein. Table 3. Sequence

SEQ ID NO: 02 - Alpha-synuclein isoform NACP140 [Homo sapiens] NCBI Reference Sequence: NP_000336.1

Aβ immunogens :

Alpha - synuclein immunogen

DAEFRHDRRX ₁ PDNEAYEXXC (SEQ ID NO: 112), wherein X ₁ is optional, and if present, X ₁ is D, and wherein XX and C independently are optional, and if present, XX can be GG , AA, KK, SS, GAGA, AGAG or KGKG.

Figure 1 shows the results of experiments comparing the titers of guinea pig sera against the immunogens DAEFRHDRRPDNEAYEGGC (SEQ ID NO: 110) and DAEFRHDRRDPDDNEAYEGGC (SEQ ID NO: 111). All immunogens contained a C-terminal GG linker and cysteine for coupling to the maleimide-activated CRM197 carrier. A squalene-based oil-in-water nanoemulsion of AddaVax containing QS21 was used as an adjuvant.

Figure 2 shows the results of experiments to measure the titers of murine sera against the immunogens DAEFRHDRRPDNEAYEGGC (SEQ ID NO: 110) and DAEFRHDRRDPDDNEAYEGGC (SEQ ID NO: 111). The peptide was coupled to the maleimide-activated CRM197 carrier via the N-terminal cysteine. QS21 in PBS was used as adjuvant.

Figure 3 shows the results of experiments to measure the titers of guinea pig sera against the immunogens DAEFRHDRRPDNEAYEGGC (SEQ ID NO: 110) and DAEFRHDRRDPDDNEAYEGGC (SEQ ID NO: 111). The peptide was coupled to the maleimide-activated CRM197 carrier via the N-terminal cysteine. QS21 in PBS was used as adjuvant.

             <![CDATA[<110> Neotope Neuroscience Limited]]> <![CDATA[<120> Polyepitope Vaccine for Alzheimer's Disease]]> <![CDATA[<130> 20-1084- WO (768-WO)]]> <![CDATA[<140> TW 110129344]]> <![CDATA[<141> 2021-08-09]]> <![CDATA[<150> US 63/080,619 ]]> <![CDATA[<151> 2020-09-18]]> <![CDATA[<160> 178 ]]> <![CDATA[<170> PatentIn version 3.5]]> <![CDATA[ <210> 1]]> <![CDATA[<211> 42]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[< 220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 1]]> Asp Ala Glu Phe Arg His Asp Ser Gly Tyr Glu Val His His Gln Lys 1 5 10 15 Val Leu Phe Phe Ala Glu Asp Val Gly Ser Asn Lys Gly Ala Ile Ile 20 25 30 Gly Leu Met Val Gly Gly Val Val Ile Ala 35 40 <![CDATA[<210> 2]]> <![CDATA[<211 > 140]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthesis Peptide]]> <![CDATA[<400> 2]]> Met Asp Val Phe Met Lys Gly Leu Ser Lys Ala Lys Glu Gly Val Val 1 5 10 15 Ala Ala Ala Glu Lys Thr Lys Gln Gly Val Ala Glu Ala Ala Gly Lys 20 25 30 Thr Lys Glu Gly Val Leu Tyr Val Gly Ser Lys Thr Lys Glu Gly Val 35 40 45 Val His Gly Val Ala Thr Val Ala Glu Lys Thr Lys Glu Gln Val Thr 50 55 60 Asn Val Gly Gly Ala Val Val Thr Gly Val Thr Ala Val Ala Gln Lys 65 70 75 80 Thr Val Glu Gly Ala Gly Ser Ile Ala Ala Ala Thr Gly Phe Val Lys 85 90 95 Lys Asp Gln Leu Gly Lys Asn Glu Glu Gly Ala Pro Gln Glu Gly Ile 100 105 110 Leu Glu Asp Met Pro Val Asp Pro Asp Asn Glu Ala Tyr Glu Met Pro 115 120 125 Ser Glu Glu Gly Tyr Gln Asp Tyr Glu Pro Glu Ala 130 135 140 <![CDATA[<210> 3]]> <![CDATA[<211> 10]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 3]]> Asp Ala Glu Phe Arg His Asp Ser Gly Tyr 1 5 10 <![CDATA[<210> 4]]> <![CDATA[<211> 9]]> <![CDATA[<212> PRT]]> <! [CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 4]]> Asp Ala Glu Phe Arg His Asp Ser Gly 1 5 <![CDATA[<210> 5]]> <![CDATA[<211> 8]]> <![CDATA[<212> PRT]]> <![CDATA[< 213>Artificial Sequence]] > <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 5]]> Asp Ala Glu Phe Arg His Asp Ser 1 5 <![ CDATA[<210> 6]]> <![CDATA[<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA [<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 6]]> Asp Ala Glu Phe Arg His Asp 1 5 <![CDATA[<210> 7 ]]> <![CDATA[<211> 6]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]] > <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 7]]> Asp Ala Glu Phe Arg His 1 5 <![CDATA[<210> 8]]> <![CDATA [<211> 5]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[< 223> Synthetic Peptides]]> <![CDATA[<400> 8]]> Asp Ala Glu Phe Arg 1 5 <![CDATA[<210> 9]]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> < ![CDATA[<400> 9]]> Asp Ala Glu Phe 1 <![CDATA[<210> 10]]> <![CDATA[<211> 3]]> <![CDATA[<212> PRT] ]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 10]] > Asp Ala Glu 1 <![CDATA[<210> 11]]> <![CDATA[<211> 9]]> <![CDATA[<212> PRT ]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 11] ]> Ala Glu Phe Arg His Asp Ser Gly Tyr 1 5 <![CDATA[<210> 12]]> <![CDATA[<211> 8]]> <![CDATA[<212> PRT]]> < ![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 12]]> Ala Glu Phe Arg His Asp Ser Gly 1 5 <![CDATA[<210> 13]]> <![CDATA[<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[< 213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 13]]> Ala Glu Phe Arg His Asp Ser 1 5 <![CDATA[<210> 14]]> <![CDATA[<211> 6]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]] > <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 14]]> Ala Glu Phe Arg His Asp 1 5 <![CDATA[ <210> 15]]> <![CDATA[<211> 5]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[< 220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 15]]> Ala Glu Phe Arg His 1 5 <![CDATA[<210> 16]]> < ![CDATA[<211> 4]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![ CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 16]]> Ala Glu Phe Arg 1 <![CDATA[<210> 17]]> <![CDATA[<211> 3]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> < ![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 17]]> Ala Glu Phe 1 <![CDATA[<210> 18]] > <![CDATA[<211> 8]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> < ![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 18]]> Glu Phe Arg His Asp Ser Gly Tyr 1 5 <![CDATA[<210> 19]]> <![CDATA [<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[< 223> Synthetic Peptides]]> <![CDATA[<400> 19]]> Glu Phe Arg His Asp Ser Gly 1 5 <![CDATA[<210> 20]]> <![CDATA[<211> 6] ]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]] > <![CDATA[<400> 20]]> Glu Phe Arg His Asp Ser 1 5 <![CDATA[<210> 21]]> <![CDATA[<211> 5]]> <![CDATA[ <212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[< 400> 21]]> Glu Phe Arg His Asp 1 5 <![CDATA[<210> 22]]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT]]> < ![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![C DATA[<400> 22]]> Glu Phe Arg His 1 <![CDATA[<210> 23]]> <![CDATA[<211> 3]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 23]]> Glu Phe Arg 1 <![CDATA[<210> 24]]> <![CDATA[<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence] ]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 24]]> Phe Arg His Asp Ser Gly Tyr 1 5 <![ CDATA[<210> 25]]> <![CDATA[<211> 6]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA [<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 25]]> Phe Arg His Asp Ser Gly 1 5 <![CDATA[<210> 26] ]> <![CDATA[<211> 5]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 26]]> Phe Arg His Asp Ser 1 5 <![CDATA[<210> 27]]> <![CDATA[< 211> 4]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 27]]> Phe Arg His Asp 1 <![CDATA[<210> 28]]> <![CDATA[<211> 3]]> <![CDATA [<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[< 223> Synthetic Peptides]]> <![CDATA[<400> 28]]> Phe Arg His 1 <![CDATA[<210> 29]]> <![CDATA[<211> 6]]> <![ CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA [<400> 29]]> Arg His Asp Ser Gly Tyr 1 5 <![CDATA[<210> 30]]> <![CDATA[<211> 5]]> <![CDATA[<212> PRT] ]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 30]] > Arg His Asp Ser Gly 1 5 <![CDATA[<210> 31]]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT]]> <![CDATA[< 213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 31]]> Arg His Asp Ser 1 <! [CDATA[<210> 32]]> <![CDATA[<211> 3]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![ CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 32]]> Arg His Asp 1 <![CDATA[<210> 33]]> < ![CDATA[<211> 5]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![ CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 33]]> His Asp Ser Gly Tyr 1 5 <![CDATA[<210> 34]]> <![CDATA[<211> 4 ]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDA TA[<223> Synthetic Peptide]]> <![CDATA[<400> 34]]> His Asp Ser Gly 1 <![CDATA[<210> 35]]> <![CDATA[<211> 3]] > <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 35]]> His Asp Ser 1 <![CDATA[<210> 36]]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT] ]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 36]] > Asp Ser Gly Tyr 1 <![CDATA[<210> 37]]> <![CDATA[<211> 3]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 37]]> Asp Ser Gly 1 <![CDATA[ <210> 38]]> <![CDATA[<211> 3]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[< 220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 38]]> Ser Gly Tyr 1 <![CDATA[<210> 39]]> <![CDATA [<211> 10]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[< 223> Synthetic Peptides]]> <![CDATA[<400> 39]]> Val Asp Pro Asp Asn Glu Ala Tyr Glu Met 1 5 10 <![CDATA[<210> 40]]> <![CDATA[< 211> 9]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]] > <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 40]]> Val Asp Pro Asp Asn Glu Ala Tyr Glu 1 5 <![CDATA[<210> 41]]> < ![CDATA[<211> 8]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![ CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 41]]> Val Asp Pro Asp Asn Glu Ala Tyr 1 5 <![CDATA[<210> 42]]> <![CDATA[< 211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 42]]> Val Asp Pro Asp Asn Glu Ala 1 5 <![CDATA[<210> 43]]> <![CDATA[<211> 6]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> < ![CDATA[<400> 43]]> Val Asp Pro Asp Asn Glu 1 5 <![CDATA[<210> 44]]> <![CDATA[<211> 5]]> <![CDATA[<212 > PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 44]]> Val Asp Pro Asp Asn 1 5 <![CDATA[<210> 45]]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT]]> <![ CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 45]]> Val Asp Pro Asp 1 <![CDATA[<210> 46]]> <![CDATA[<211> 3]]> < ![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <! [CDATA[<400> 46]]> Val Asp Pro 1 <![CDATA[<210> 47]]> <![CDATA[<211> 9]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 47]]> Asp Pro Asp Asn Glu Ala Tyr Glu Met 1 5 <![CDATA[<210> 48]]> <![CDATA[<211> 8]]> <![CDATA[<212> PRT]]> <![CDATA [<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 48]]> Asp Pro Asp Asn Glu Ala Tyr Glu 1 5 <![CDATA[<210> 49]]> <![CDATA[<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 49]]> Asp Pro Asp Asn Glu Ala Tyr 1 5 < ![CDATA[<210> 50]]> <![CDATA[<211> 6]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <! [CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 50]]> Asp Pro Asp Asn Glu Ala 1 5 <![CDATA[<210> 51]]> <![CDATA[<211> 5]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>] ]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 51]]> Asp Pro As p Asn Glu 1 5 <![CDATA[<210> 52]]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 52]]> Asp Pro Asp Asn 1 <![CDATA[ <210> 53]]> <![CDATA[<211> 3]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[< 220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 53]]> Asp Pro Asp 1 <![CDATA[<210> 54]]> <![CDATA [<211> 8]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[< 223> Synthetic Peptides]]> <![CDATA[<400> 54]]> Pro Asp Asn Glu Ala Tyr Glu Met 1 5 <![CDATA[<210> 55]]> <![CDATA[<211> 7 ]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide] ]> <![CDATA[<400> 55]]> Pro Asp Asn Glu Ala Tyr Glu 1 5 <![CDATA[<210> 56]]> <![CDATA[<211> 6]]> <![ CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA [<400> 56]]> Pro Asp Asn Glu Ala Tyr 1 5 <![CDATA[<210> 57]]> <![CDATA[<211> 5]]> <![CDATA[<212> PRT] ]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide] ]> <![CDATA[<400> 57]]> Pro Asp Asn Glu Ala 1 5 <![CDATA[<210> 58]]> <![CDATA[<211> 4]]> <![CDATA[ <212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[< 400> 58]]> Pro Asp Asn Glu 1 <![CDATA[<210> 59]]> <![CDATA[<211> 3]]> <![CDATA[<212> PRT]]> <![ CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 59]]> Pro Asp Asn 1 <![CDATA[<210> 60]]> <![CDATA[<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> < ![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 60]]> Asp Asn Glu Ala Tyr Glu Met 1 5 <![CDATA[< 210> 61]]> <![CDATA[<211> 6]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220 >]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 61]]> Asp Asn Glu Ala Tyr Glu 1 5 <![CDATA[<210> 62]]> < ![CDATA[<211> 5]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![ CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 62]]> Asp Asn Glu Ala Tyr 1 5 <![CDATA[<210> 63]]> <![CDATA[<211> 4 ]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>] ]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 63]]> Asp Asn Glu Ala 1 <![CDATA[<210> 64]]> <![CDATA[< 211> 3]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 64]]> Asp Asn Glu 1 <![CDATA[<210> 65]]> <![CDATA[<211> 6]]> <![CDATA[ <212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[< 400> 65]]> Asn Glu Ala Tyr Glu Met 1 5 <![CDATA[<210> 66]]> <![CDATA[<211> 5]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 66]]> Asn Glu Ala Tyr Glu 1 5 <![CDATA[<210> 67]]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 67]]> Asn Glu Ala Tyr 1 <![CDATA [<210> 68]]> <![CDATA[<211> 3]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[ <220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 68]]> Asn Glu Ala 1 <![CDATA[<210> 69]]> <![ CDATA[<211> 5]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 69]]> Glu Ala Tyr Glu Met 1 5 <![CDATA[<210> 70]]> <![CDATA[< 211> 4]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 70]]> Glu Ala Tyr Glu 1 <![CDATA[<210> 71]]> <![CDATA[<211> 3]]> <![CDATA [<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[ <400> 71]]> Glu Ala Tyr 1 <![CDATA[<210> 72]]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT]]> <![ CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 72]]> Ala Tyr Glu Met 1 <![CDATA[<210> 73]]> <![CDATA[<211> 3]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 73]]> Ala Tyr Glu 1 <![CDATA[<210> 74] ]> <![CDATA[<211> 3]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 74]]> Tyr Glu Met 1 <![CDATA[<210> 75]]> <![CDATA[<211> 10 ]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 75]]> Ala Thr Gly Phe Val Lys Lys Asp Gln Leu 1 5 10 <![CDATA[<210> 76]]> <![CDATA[<211> 9]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide ]]> <![CDATA[<400> 76]]> Ala Thr Gly Phe Val Lys Lys Asp Gln 1 5 <![CDATA[<210> 77]]> <![CDATA[<211> 8]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> < ![CDATA[<400> 77]]> Ala Thr Gly Phe Val Lys Lys Asp 1 5 <![CDATA[<210> 78]]> <![CDATA[<211> 7]]> <![CDATA[ <212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[< 400> 78]]> Ala Thr Gly Phe Val Lys Lys 1 5 <![CDATA[<210> 79]]> <![CDATA[<211> 6]]> <![CDATA[<212> PRT]] > <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 79]]> Ala Thr Gly Phe Val Lys 1 5 <![CDATA[<210> 80]]> <![CDATA[<211> 5]]> <![CDATA[<212> PRT]]> <![CDATA[< 213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 80]]> Ala Thr Gly Phe Val 1 5 <![CDATA[<210> 81]]> <![CDATA [<211> 4]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[< 223> Synthetic Peptides]]> <![ CDATA[<400> 81]]> Ala Thr Gly Phe 1 <![CDATA[<210> 82]]> <![CDATA[<211> 3]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 82]]> Ala Thr Gly 1 <![CDATA[<210> 83]]> <![CDATA[<211> 9]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence] ]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 83]]> Thr Gly Phe Val Lys Lys Asp Gln Leu 1 5 < ![CDATA[<210> 84]]> <![CDATA[<211> 8]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <! [CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 84]]> Thr Gly Phe Val Lys Lys Asp Gln 1 5 <![CDATA[< 210> 85]]> <![CDATA[<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220 >]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 85]]> Thr Gly Phe Val Lys Lys Asp 1 5 <![CDATA[<210> 86]]> <![CDATA[<211> 6]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <! [CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 86]]> Thr Gly Phe Val Lys Lys 1 5 <![CDATA[<210> 87]]> <![CDATA[<211 > 5]]> <![CDATA[<212> PRT]]> <![CDATA[<213> people Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 87]]> Thr Gly Phe Val Lys 1 5 <! [CDATA[<210> 88]]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![ CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 88]]> Thr Gly Phe Val 1 <![CDATA[<210> 89]]> <![CDATA[<211> 3]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <! [CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 89]]> Thr Gly Phe 1 <![CDATA[<210> 90]]> <![CDATA[<211> 8]] > <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 90]]> Gly Phe Val Lys Lys Asp Gln Leu 1 5 <![CDATA[<210> 91]]> <![CDATA[<211> 7]]> <![CDATA [<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[ <400> 91]]> Gly Phe Val Lys Lys Asp Gln 1 5 <![CDATA[<210> 92]]> <![CDATA[<211> 6]]> <![CDATA[<212> PRT] ]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 92]] > Gly Phe Val Lys Lys Asp 1 5 <![CDATA[<210> 93]]> <![CDATA[<211> 5]]> <![C DATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA [<400> 93]]> Gly Phe Val Lys Lys 1 5 <![CDATA[<210> 94]]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT]] > <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 94]]> Gly Phe Val Lys 1 <![CDATA[<210> 95]]> <![CDATA[<211> 3]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 95]]> Gly Phe Val 1 <![CDATA[< 210> 96]]> <![CDATA[<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220 >]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 96]]> Phe Val Lys Lys Asp Gln Leu 1 5 <![CDATA[<210> 97]]> <![CDATA[<211> 6]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <! [CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 97]]> Phe Val Lys Lys Asp Gln 1 5 <![CDATA[<210> 98]]> <![CDATA[<211 > 5]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthesis Peptide]]> <![CDATA[<400> 98]]> Phe Val Lys Lys Asp 1 5 <![CDATA[<210> 99]]> <! [CDATA[<211> 4]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA [<223> Synthetic Peptides]]> <![CDATA[<400> 99]]> Phe Val Lys Lys 1 <![CDATA[<210> 100]]> <![CDATA[<211> 3]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> < ![CDATA[<400> 100]]> Phe Val Lys 1 <![CDATA[<210> 101]]> <![CDATA[<211> 6]]> <![CDATA[<212> PRT]] > <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 101]]> Val Lys Lys Asp Gln Leu 1 5 <![CDATA[<210> 102]]> <![CDATA[<211> 5]]> <![CDATA[<212> PRT]]> <![CDATA[< 213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 102]]> Val Lys Lys Asp Gln 1 5 <![CDATA[<210> 103]]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> < ![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 103]]> Val Lys Lys Asp 1 <![CDATA[<210> 104] ]> <![CDATA[<211> 3]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 104]]> Val Lys Lys 1 <![CDATA[<210> 105]]> <![CDATA[<211> 5]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>] ]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 105]]> Lys Lys Asp Gln Leu 1 5 <![CDATA[<210> 106]]> <![CDATA [<211> 4]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[< 223> Synthetic Peptides]]> <![CDATA[<400> 106]]> Lys Lys Asp Gln 1 <![CDATA[<210> 107]]> <![CDATA[<211> 3]]> <! [CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![ CDATA[<400> 107]]> Lys Lys Asp 1 <![CDATA[<210> 108]]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT]]> < ![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 108]]> Lys Asp Gln Leu 1 <![CDATA[<210> 109]]> <![CDATA[<211> 3]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence] ]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 109]]> Lys Asp Gln 1 <![CDATA[<210> 110]]> <![CDATA[<211> 19]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>] ]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 110]]> Asp Ala Glu Phe Arg His Asp Arg Arg Pro Asp Asn Glu Ala Tyr Glu 1 5 10 15 Gly Gly Cys <![CDATA[<210> 111]]> <![CDATA[<211> 20]]> <![CDATA[<212> PRT]] > <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 111]]> Asp Ala Glu Phe Arg His Asp Arg Arg Asp Pro Asp Asn Glu Ala Tyr 1 5 10 15 Glu Gly Gly Cys 20 <![CDATA[<210> 112]]> <![CDATA[<211> 20]]> < ![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <! [CDATA[<220>]]> <![CDATA[<221> MISC_FEATURE]]> <![CDATA[<222> (10)..(10)]]> <![CDATA[<223> as appropriate exists, or D if it exists]]> <![CDATA[<220>]]> <![CDATA[<221> MISC_FEATURE]]> <![CDATA[<222> (18)..(19)] ]> <![CDATA[<223> If present, G, A, K, or S if present]]> <![CDATA[<400> 112]]> Asp Ala Glu Phe Arg His Asp Arg Arg Xaa Pro Asp Asn Glu Ala Tyr 1 5 10 15 Glu Xaa Xaa Cys 20 <![CDATA[<210> 113]]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT]] > <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 113]]> Arg Val Arg Arg 1 <![CDATA[<210> 114]]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT]]> <![CD ATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 114]]> Gly Ala Gly Ala 1 <![CDATA[<210> 115]]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 115]]> Ala Gly Ala Gly 1 <![CDATA[<210> 116 ]]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]] > <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 116]]> Lys Gly Lys Gly 1 <![CDATA[<210> 117]]> <![CDATA[<211 > 9]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthesis Peptide]]> <![CDATA[<400> 117]]> Ala Glu Phe Arg His Asp Ser Gly Cys 1 5 <![CDATA[<210> 118]]> <![CDATA[<211> 8]] > <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 118]]> Asp Ala Glu Phe Arg His Asp Cys 1 5 <![CDATA[<210> 119]]> <![CDATA[<211> 8]]> <![CDATA [<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[ <400> 119]]> Cys Pro Asp Asn Glu Ala Tyr Glu 1 5 <![CDATA[ <210> 120]]> <![CDATA[<211> 8]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[< 220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 120]]> Asp Pro Asp Asn Glu Ala Tyr Cys 1 5 <![CDATA[<210> 121] ]> <![CDATA[<211> 10]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 121]]> Val His His Gln Lys Leu Val Phe Phe Ala 1 5 10 <![CDATA[<210> 122]]> <![CDATA[<211> 9]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <! [CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 122]]> Val His His Gln Lys Leu Val Phe Phe 1 5 <![CDATA[<210> 123]]> <![CDATA [<211> 8]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[< 223> Synthetic Peptides]]> <![CDATA[<400> 123]]> Val His His Gln Lys Leu Val Phe 1 5 <![CDATA[<210> 124]]> <![CDATA[<211> 7 ]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide] ]> <![CDATA[<400> 124]]> Val His His Gln Lys Leu Val 1 5 <![CDATA[<210> 125]]> <![CDATA[<211> 6]]> <![ CDATA[<212> PRT]]> <![CDATA [<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 125]]> Val His His Gln Lys Leu 1 5 <![CDATA[<210> 126]]> <![CDATA[<211> 10]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence] ]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 126]]> His His Gln Lys Leu Val Phe Phe Ala Glu 1 5 10 <![CDATA[<210> 127]]> <![CDATA[<211> 9]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 127]]> His His Gln Lys Leu Val Phe Phe Ala 1 5 <![ CDATA[<210> 128]]> <![CDATA[<211> 8]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA [<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 128]]> His His Gln Lys Leu Val Phe Phe 1 5 <![CDATA[<210> 129]]> <![CDATA[<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>] ]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 129]]> His His Gln Lys Leu Val Phe 1 5 <![CDATA[<210> 130]]> <! [CDATA[<211> 6]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA [<223> Synthetic Peptides]]> <![CDATA[<400> 1 30]]> His His Gln Lys Leu Val 1 5 <![CDATA[<210> 131]]> <![CDATA[<211> 5]]> <![CDATA[<212> PRT]]> <! [CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 131]]> His His Gln Lys Leu 1 5 <![CDATA[<210> 132]]> <![CDATA[<211> 10]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial sequence ]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 132]]> His Gln Lys Leu Val Phe Phe Ala Glu Asp 1 5 10 <![CDATA[<210> 133]]> <![CDATA[<211> 9]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]] > <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 133]]> His Gln Lys Leu Val Phe Phe Ala Glu 1 5 <! [CDATA[<210> 134]]> <![CDATA[<211> 8]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![ CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 134]]> His Gln Lys Leu Val Phe Phe Ala 1 5 <![CDATA[<210 > 135]]> <![CDATA[<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220> ]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 135]]> His Gln Lys Leu Val Phe Phe 1 5 <![CDATA[<210> 136]]> < ![CDATA[<211> 6]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> < ![CDATA[<400> 136]]> His Gln Lys Leu Val Phe 1 5 <![CDATA[<210> 137]]> <![CDATA[<211> 5]]> <![CDATA[<212 > PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 137]]> His Gln Lys Leu Val 1 5 <![CDATA[<210> 138]]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT]]> <![ CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 138]]> His Gln Lys Leu 1 <![CDATA[<210> 139]]> <![CDATA[<211> 10]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 139]]> Gln Lys Leu Val Phe Phe Ala Glu Asp Val 1 5 10 < ![CDATA[<210> 140]]> <![CDATA[<211> 9]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <! [CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 140]]> Gln Lys Leu Val Phe Phe Ala Glu Asp 1 5 <![CDATA[ <210> 141]]> <![CDATA[<211> 8]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[< 220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<4 00> 141]]> Gln Lys Leu Val Phe Phe Ala Glu 1 5 <![CDATA[<210> 142]]> <![CDATA[<211> 7]]> <![CDATA[<212> PRT] ]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 142]] > Gln Lys Leu Val Phe Phe Ala 1 5 <![CDATA[<210> 143]]> <![CDATA[<211> 6]]> <![CDATA[<212> PRT]]> <![CDATA [<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 143]]> Gln Lys Leu Val Phe Phe 1 5 <![CDATA[<210> 144]]> <![CDATA[<211> 5]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence] ]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 144]]> Gln Lys Leu Val Phe 1 5 <![CDATA[ <210> 145]]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[< 220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 145]]> Gln Lys Leu Val 1 <![CDATA[<210> 146]]> <![ CDATA[<211> 3]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[ <223> Synthetic Peptides]]> <![CDATA[<400> 146]]> Gln Lys Leu 1 <![CDATA[<210> 147]]> <![CDATA[<211> 10]]> <! [CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![C DATA[<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 147]]> Lys Leu Val Phe Phe Ala Glu Asp Val Gly 1 5 10 <![CDATA [<210> 148]]> <![CDATA[<211> 9]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[ <220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 148]]> Lys Leu Val Phe Phe Ala Glu Asp Val 1 5 <![CDATA[<210> 149]]> <![CDATA[<211> 8]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>] ]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 149]]> Lys Leu Val Phe Phe Ala Glu Asp 1 5 <![CDATA[<210> 150]]> < ![CDATA[<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![ CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 150]]> Lys Leu Val Phe Phe Ala Glu 1 5 <![CDATA[<210> 151]]> <![CDATA[<211 > 6]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthesis Peptides]]> <![CDATA[<400> 151]]> Lys Leu Val Phe Phe Ala 1 5 <![CDATA[<210> 152]]> <![CDATA[<211> 5]]> <! [CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![ CDATA[<400> 152]]> Lys Leu Val Ph e Phe 1 5 <![CDATA[<210> 153]]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial sequence ]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 153]]> Lys Leu Val Phe 1 <![CDATA[< 210> 154]]> <![CDATA[<211> 3]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220 >]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 154]]> Lys Leu Val 1 <![CDATA[<210> 155]]> <![CDATA[ <211> 9]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[<223 > Synthetic Peptides]]> <![CDATA[<400> 155]]> Leu Val Phe Phe Ala Glu Asp Val Gly 1 5 <![CDATA[<210> 156]]> <![CDATA[<211> 8 ]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide] ]> <![CDATA[<400> 156]]> Leu Val Phe Phe Ala Glu Asp Val 1 5 <![CDATA[<210> 157]]> <![CDATA[<211> 7]]> <! [CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![ CDATA[<400> 157]]> Leu Val Phe Phe Ala Glu Asp 1 5 <![CDATA[<210> 158]]> <![CDATA[<211> 6]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 158]]> Leu Val Phe Phe Ala Glu 1 5 <![CDATA[<210> 159]]> <![CDATA[ <211> 5]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[<223 > Synthetic Peptides]]> <![CDATA[<400> 159]]> Leu Val Phe Phe Ala 1 5 <![CDATA[<210> 160]]> <![CDATA[<211> 4]]> < ![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <! [CDATA[<400> 160]]> Leu Val Phe Phe 1 <![CDATA[<210> 161]]> <![CDATA[<211> 3]]> <![CDATA[<212> PRT]] > <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 161]]> Leu Val Phe 1 <![CDATA[<210> 162]]> <![CDATA[<211> 8]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial sequence ]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 162]]> Val Phe Phe Ala Glu Asp Val Gly 1 5 < ![CDATA[<210> 163]]> <![CDATA[<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <! [CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 163]]> Val Phe Phe Ala Glu Asp Val 1 5 <![CDATA[<210 > 164]]> <![CDATA[<211> 6]]> <![CDATA[<212> PR T]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![ CDATA[<400> 164]]> Val Phe Phe Ala Glu Asp 1 5 <![CDATA[<210> 165]]> <![CDATA[<211> 5]]> <![CDATA[<212> PRT ]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 165] ]> Val Phe Phe Ala Glu 1 5 <![CDATA[<210> 166]]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT]]> <![CDATA[ <213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 166]]> Val Phe Phe Ala 1 < ![CDATA[<210> 167]]> <![CDATA[<211> 3]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <! [CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 167]]> Val Phe Phe 1 <![CDATA[<210> 168]]> <![CDATA[<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <! [CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 168]]> Phe Phe Ala Glu Asp Val Gly 1 5 <![CDATA[<210> 169]]> <![CDATA[< 211> 6]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 169]]> Phe Phe Ala Glu Asp Val 1 5 <![CDATA[<210> 170]]> <![CDATA[<211> 5]]> < ![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CD ATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 170]]> Phe Phe Ala Glu Asp 1 5 <![CDATA[<210> 171] ]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 171]]> Phe Phe Ala Glu 1 <![CDATA[<210> 172]]> <![CDATA[<211> 3]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide ]]> <![CDATA[<400> 172]]> Phe Phe Ala 1 <![CDATA[<210> 173]]> <![CDATA[<211> 6]]> <![CDATA[<212 > PRT]]> <![CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 173]]> Phe Ala Glu Asp Val Gly 1 5 <![CDATA[<210> 174]]> <![CDATA[<211> 5]]> <![CDATA[<212> PRT]]> <! [CDATA[<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 174]]> Phe Ala Glu Asp Val 1 5 <![CDATA[<210> 175]]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Artificial sequence ]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptides]]> <![CDATA[<400> 175]]> Phe Ala Glu Asp 1 <![CDATA[< 210> 176]]> <![CDATA[<211> 3]]> <![CDATA[<212> PRT]]> <![CDATA [<213> Artificial Sequence]]> <![CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 176]]> Phe Ala Glu 1 < ![CDATA[<210> 177]]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <! [CDATA[<220>]]> <![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 177]]> Gly Gly Gly Ser 1 <![CDATA[<210> 178]] > <![CDATA[<211> 5]]> <![CDATA[<212> PRT]]> <![CDATA[<213> Manual Sequence]]> <![CDATA[<220>]]> < ![CDATA[<223> Synthetic Peptide]]> <![CDATA[<400> 178]]> Gly Gly Gly Gly Ser 1 5
      

Claims (67)

A polypeptide comprising a first peptide comprising 3 to 10 amino acids from residues 1 to 10 or 12 to 25 of SEQ ID NO:01 linked to a second peptide comprising 3 to 10 amino acids from residues 81 to 140 of SEQ ID NO:02. The polypeptide of claim 1, wherein the second peptide is derived from the C-terminal region of alpha-synuclein (residues 111 to 131 of SEQ ID NO:02). The polypeptide of claim 1, wherein the first peptide is located at the N-terminus of the second peptide. The polypeptide of claim 1, wherein the first peptide is C-terminal to the second peptide. The polypeptide of any one of claims 1 to 4, wherein: (a) The first peptide comprises an amino acid sequence selected from the group consisting of: DAEFRHDSGY (SEQ ID NO: 03), DAEFRHDSG (SEQ ID NO: 04), DAEFRHDS (SEQ ID NO: 05), DAEFRHD (SEQ ID NO: 06), DAEFRH (SEQ ID NO: 07), DAEFR (SEQ ID NO: 08), DAEF (SEQ ID NO: 09), DAE (SEQ ID NO: 10), AEFRHDSGY (SEQ ID NO: 11), AEFRHDSG (SEQ ID NO: 12), AEFRHDS (SEQ ID NO: 13), AEFRHD (SEQ ID NO: 14), AEFRH (SEQ ID NO: 15), AEFR (SEQ ID NO: 16), AEF (SEQ ID NO: 17), EFRHDSGY (SEQ ID NO: 18), EFRHDSG (SEQ ID NO: 19), EFRHDS (SEQ ID NO: 20), EFRHD (SEQ ID NO: 21), EFRH (SEQ ID NO: 22), EFR (SEQ ID NO: 23), FRHDSGY (SEQ ID NO: 24), FRHDSG (SEQ ID NO: 25), FRHDS (SEQ ID NO: 26), FRHD (SEQ ID NO: 27), FRH (SEQ ID NO: 28), RHDSGY (SEQ ID NO: 29), RHDSG (SEQ ID NO: 30), RHDS (SEQ ID NO: 31), RHD (SEQ ID NO: 32), HDSGY (SEQ ID NO: 33), HDSG (SEQ ID NO: 34), HDS (SEQ ID NO: 35), DSGY (SEQ ID NO: 36), DSG (SEQ ID NO: 37), SGY (SEQ ID NO: 38), VHHQKLVFFA (SEQ ID NO: 121), VHHQKLVFF (SEQ ID NO: 122), VHHQKLVF (SEQ ID NO: 123), VHHQKLV (SEQ ID NO: 124), VHHQKL (SEQ ID NO: 125), HHQKLVFFAE (SEQ ID NO: 126), HHQKLVFFA (SEQ ID NO: 127), HHQKLVFF (SEQ ID NO: 128), HHQKLVF (SEQ ID NO: 129), HHQKLV (SEQ ID NO: 130), HHQKL (SEQ ID NO: 131), HQKLVFFAED (SEQ ID NO: 132), HQKLVFFAE (SEQ ID NO: 133), HQKLVFFA (SEQ ID NO: 134), HQKLVFF (SEQ ID NO: 135), HQKLVF (SEQ ID NO: 136), HQKLV (SEQ ID NO: 137), HQKL (SEQ ID NO: 138), QKLVFFAEDV (SEQ ID NO: 139), QKLVFFAED (SEQ ID NO: 140), QKLVFFAE (SEQ ID NO: 141), QKLVFFA (SEQ ID NO: 142), QKLVFF (SEQ ID NO: 143), QKLVF (SEQ ID NO: 144), QKLV (SEQ ID NO: 145), QKL (SEQ ID NO: 146), KLVFFAEDVG (SEQ ID NO: 147), KLVFFAEDV (SEQ ID NO: 148), KLVFFAED (SEQ ID NO: 149), KLVFFAE (SEQ ID NO: 150), KLVFFA (SEQ ID NO: 151), KLVFF (SEQ ID NO: 152), KLVF (SEQ ID NO: 153), KLV (SEQ ID NO: 154), LVFFAEDVG (SEQ ID NO: 155), LVFFAEDV (SEQ ID NO: 156), LVFFAED (SEQ ID NO: 157), LVFFAE (SEQ ID NO: 158), LVFFA (SEQ ID NO: 159), LVFF (SEQ ID NO: 160), LVF (SEQ ID NO: 161), VFFAEDVG (SEQ ID NO: 162), VFFAEDV (SEQ ID NO: 163), VFFAED (SEQ ID NO: 164), VFFAE (SEQ ID NO: 165), VFFA (SEQ ID NO: 166), VFF (SEQ ID NO: 167), FFAEDVG (SEQ ID NO: 168), FFAEDV (SEQ ID NO: 169), FFAED (SEQ ID NO: 170), FFAE (SEQ ID NO: 171), FFA (SEQ ID NO: 172), FAEDVG (SEQ ID NO: 173), FAEDV (SEQ ID NO: 174), FAED (SEQ ID NO: 175), FAE (SEQ ID NO: 176); and (b) the second peptide comprises an amino acid sequence selected from the group consisting of: VDPDNEAYEM (SEQ ID NO: 39), VDPDNEAYE (SEQ ID NO: 40), VDPDNEAY (SEQ ID NO: 41), VDPDNEA (SEQ ID NO: 42), VDPDNE (SEQ ID NO: 43), VDPDN (SEQ ID NO: 44), VDPD (SEQ ID NO: 45), VDP (SEQ ID NO: 46), DPDNEAYEM (SEQ ID NO: 47), DPDNEAYE (SEQ ID NO: 48), DPDNEAY (SEQ ID NO: 49), DPDNEA (SEQ ID NO: 50), DPDNE (SEQ ID NO: 51), DPDN (SEQ ID NO: 52), DPD (SEQ ID NO: 53), PDNEAYEM (SEQ ID NO: 54), PDNEAYE (SEQ ID NO: 55), PDNEAY (SEQ ID NO: 56), PDNEA (SEQ ID NO: 57), PDNE (SEQ ID NO: 58), PDN (SEQ ID NO: 59), DNEAYEM (SEQ ID NO: 60), DNEAYE (SEQ ID NO: 61), DNEAY (SEQ ID NO: 62), DNEA (SEQ ID NO: 63), DNE (SEQ ID NO: 64), NEAYEM (SEQ ID NO: 65), NEAYE (SEQ ID NO: 66), NEAY (SEQ ID NO: 67), NEA (SEQ ID NO: 68), EAYEM (SEQ ID NO: 69), EAYE (SEQ ID NO: 70), EAY (SEQ ID NO: 71), AYEM (SEQ ID NO: 72), AYE (SEQ ID NO: 73), YEM (SEQ ID NO: 74), ATGFVKKDQL (SEQ ID NO:75), ATGFVKKDQ (SEQ ID NO: 76), ATGFVKKD (SEQ ID NO: 77), ATGFVKK (SEQ ID NO:78), ATGFVK (SEQ ID NO:79), ATGFV (SEQ ID NO:80), ATGF (SEQ ID NO:81), ATG (SEQ ID NO: 82), TGFVKKDQL (SEQ ID NO: 83), TGFVKKDQ (SEQ ID NO: 84), TGFVKKD (SEQ ID NO: 85), TGFVKK (SEQ ID NO: 86), TGFVK (SEQ ID NO:87), TGFV (SEQ ID NO: 88), TGF (SEQ ID NO:89), GFVKKDQL (SEQ ID NO: 90), GFVKKDQ (SEQ ID NO: 91), GFVKKD (SEQ ID NO: 92), GFVKK (SEQ ID NO: 93), GFVK (SEQ ID NO: 94), GFV (SEQ ID NO: 95), FVKKDQL (SEQ ID NO: 96), FVKKDQ (SEQ ID NO: 97), FVKKD (SEQ ID NO: 98) FVKK (SEQ ID NO: 99), FVK (SEQ ID NO: 100), VKKDQL (SEQ ID NO: 101), VKKDQ (SEQ ID NO: 102), VKKD (SEQ ID NO: 103), VKK (SEQ ID NO: 104), KKDQL (SEQ ID NO: 105), KKDQ (SEQ ID NO: 106), KKD (SEQ ID NO: 107), KDQL (SEQ ID NO: 108), and KDQ (SEQ ID NO: 109). The polypeptide of any one of claims 1 to 5, wherein the first peptide and the second peptide are linked by a cleavable linker. The polypeptide of claim 6, wherein the cleavable linker comprises an amino acid sequence. The polypeptide of claim 7, wherein the amino acid sequence comprises arginine-arginine (Arg-Arg), arginine-valine-arginine-arginine (Arg-Val-Arg-Arg) (SEQ ID NO: 113)), valine-citrulline (Val-Cit), valine-arginine (Val-Arg), valine-lysine (Val-Lys), valine Acid-alanine (Val-Ala), phenylalanine-lysine (Phe-Lys), glycine-alanine-glycine-alanine (Gly-Ala-Gly-Ala; SEQ ID NO: 114) , alanine-glycine-alanine-glycine (Ala-Gly-Ala-Gly; SEQ ID NO: 115) or lysine-glycine-lysine-glycine (Lys-Gly- Lys-Gly; SEQ ID NO: 116). The polypeptide of any one of claims 1 to 8, which further comprises a linker that connects the C-terminal portion of the polypeptide or the N-terminal portion of the polypeptide to a carrier. The polypeptide of claim 9, wherein the linker comprises an amino acid sequence selected from the group consisting of: GG, GGG, AA, AAA, KK, KKK, SS, SSS, GAGA (SEQ ID NO: 114), AGAG (SEQ ID NO: 115) and KGKG (SEQ ID NO: 116). The polypeptide of any one of claims 1 to 10, wherein, if present, the polypeptide or the linker to the carrier further comprises a C-terminal cysteine (C). The polypeptide of any one of claims 1 to 11, wherein the first peptide is DAEFRHD (SEQ ID NO: 06). The polypeptide of any one of claims 1 to 11, wherein the first peptide is DAEFR (SEQ ID NO:08). The polypeptide of any one of claims 1 to 11, wherein the first peptide is EFRHD (SEQ ID NO: 21). The polypeptide of any one of claims 1 to 11, wherein the second peptide comprises 5 to 10 amino acids. The polypeptide of any one of claims 1 to 11, wherein the second peptide comprises the amino acid sequence PDNEAYE (SEQ ID NO: 55). The polypeptide of any one of claims 1 to 11, wherein the second peptide comprises the amino acid sequence DPDNEAYE (SEQ ID NO: 48). The polypeptide of any one of claims 1 to 11, wherein the second peptide comprises the amino acid sequence ATGFVKK (SEQ ID NO:78), TGFVKKD (SEQ ID NO:85) or GFVKKDQ (SEQ ID NO:91). The polypeptide of claim 1, comprising the amino acid sequence of DAEFRHDRRPDNEAYEGGC (SEQ ID NO: 110). The polypeptide of claim 1, comprising the amino acid sequence DAEFRHDRRDPDDNEAYEGGC (SEQ ID NO: 111). The polypeptide of claim 1, comprising the amino acid sequence of DAEFRHDRRX ₁ PDNEAYEXXC (SEQ ID NO: 112), wherein X ₁ is optionally present, and X ₁ , if present, is D, and wherein XX and C are independently Optionally, and if present, XX can be GG, AA, KK, SS, GAGA (SEQ ID NO: 114), AGAG (SEQ ID NO: 115), or KGKG (SEQ ID NO: 116). An immunotherapy composition comprising the polypeptide of any one of claims 1 to 21, wherein the polypeptide is linked to a carrier. The immunotherapy composition of claim 22, wherein the carrier comprises serum albumin, immunoglobulin molecules, thyroglobulin, ovalbumin, tetanus toxoid (TT), diphtheria toxoid (DT), genetically modified diphtheria toxin cross-reactive substance (CRM), CRM197, meningococcal outer membrane protein complex (OMPC) and Haemophilus influenzae protein D ( H. influenzae protein D ; HiD), rEPA ( Pseudomonas aeruginosa exotoxin) A)), KLH (keyhole limpet hemocyanin) and flagellin. The immunotherapy composition of claim 23, wherein the carrier is CRM197. The immunotherapy composition of claim 23, wherein the carrier is diphtheria toxoid. A pharmaceutical formulation comprising (a) the polypeptide of any one of claims 1 to 21 or the immunotherapy composition of any one of claims 22 to 25, and (b) at least one adjuvant. The pharmaceutical formulation of claim 26, wherein the adjuvant is selected from the group consisting of: aluminum hydroxide, aluminum phosphate, aluminum sulfate, 3-de-O-acylated monophosphoryl lipid A (MPL), QS -21, TQL1055, QS-18, QS-17, QS-7, Complete Freund's Adjuvant (CFA), Incomplete Freund's Adjuvant (IFA), oil-in-water emulsions (such as squalene or peanut oil), CpG, polyglutamic acid, polylysine, AddaVax™, MF59®, and combinations thereof. The pharmaceutical formulation of claim 27, wherein the adjuvant is QS-21 or TQL1055. The pharmaceutical formulation of claim 27, wherein the adjuvant is MPL. The pharmaceutical formulation of claim 27, wherein the adjuvant is a combination of MPL and QS-21, or a combination of MPL and TQL1055. The pharmaceutical formulation of any one of claims 26 to 30, wherein the adjuvant comprises a liposomal formulation. The pharmaceutical formulation of any one of claims 26 to 31, wherein the composition comprises at least one pharmaceutically acceptable diluent. The pharmaceutical formulation of any one of claims 26 to 32, comprising a multiple antigen presentation system (MAP). The pharmaceutical formulation of claim 33, wherein the MAP comprises a Lys-based dendritic structure, a helper T cell epitope, an immunostimulatory lipophilic moiety, a cell penetrating peptide, free radical-induced polymerization, as an antigen presentation platform One or more of self-assembled nanoparticles and gold nanoparticles. An immunotherapy composition comprising a first peptide sequence comprising 3 to 10 amine groups from the first ten N-terminal residues or residues 12 to 25 of SEQ ID NO:01 and a second peptide sequence acid residues, the second peptide sequence comprises 3 to 8 amino acids from residues 81 to 140 of SEQ ID NO:02. The immunotherapy composition of claim 35, wherein: (a) the first peptide sequence comprises an amino acid sequence selected from the group consisting of: DAEFRHDSGY (SEQ ID NO: 03), DAEFRHDSG (SEQ ID NO: 04), DAEFRHDS (SEQ ID NO: 05), DAEFRHD (SEQ ID NO: 06), DAEFRH (SEQ ID NO: 07), DAEFR (SEQ ID NO: 08), DAEF (SEQ ID NO: 09), DAE (SEQ ID NO: 10), AEFRHDSGY (SEQ ID NO: 11), AEFRHDSG (SEQ ID NO: 12), AEFRHDS (SEQ ID NO: 13), AEFRHD (SEQ ID NO: 14), AEFRH (SEQ ID NO: 15), AEFR (SEQ ID NO: 16), AEF (SEQ ID NO: 17), EFRHDSGY (SEQ ID NO: 18), EFRHDSG (SEQ ID NO: 19), EFRHDS (SEQ ID NO: 20), EFRHD (SEQ ID NO: 21), EFRH (SEQ ID NO: 22), EFR (SEQ ID NO: 23), FRHDSGY (SEQ ID NO: 24), FRHDSG (SEQ ID NO: 25), FRHDS (SEQ ID NO: 26), FRHD (SEQ ID NO: 27), FRH (SEQ ID NO: 28), RHDSGY (SEQ ID NO: 29), RHDSG (SEQ ID NO: 30), RHDS (SEQ ID NO: 31), RHD (SEQ ID NO: 32), HDSGY (SEQ ID NO: 33), HDSG (SEQ ID NO: 34), HDS (SEQ ID NO: 35), DSGY (SEQ ID NO: 36), DSG (SEQ ID NO: 37), SGY (SEQ ID NO: 38), VHHQKLVFFA (SEQ ID NO: 121), VHHQKLVFF (SEQ ID NO: 122), VHHQKLVF (SEQ ID NO: 123), VHHQKLV (SEQ ID NO: 124), VHHQKL (SEQ ID NO: 125), HHQKLVFFAE (SEQ ID NO: 126), HHQKLVFFA (SEQ ID NO: 127), HHQKLVFF (SEQ ID NO: 128), HHQKLVF (SEQ ID NO: 129), HHQKLV (SEQ ID NO: 130), HHQKL (SEQ ID NO: 131), HQKLVFFAED (SEQ ID NO: 132), HQKLVFFAE (SEQ ID NO: 133), HQKLVFFA (SEQ ID NO: 134), HQKLVFF (SEQ ID NO: 135), HQKLVF (SEQ ID NO: 136), HQKLV (SEQ ID NO: 137), HQKL (SEQ ID NO: 138), QKLVFFAEDV (SEQ ID NO: 139), QKLVFFAED (SEQ ID NO: 140), QKLVFFAE (SEQ ID NO: 141), QKLVFFA (SEQ ID NO: 142), QKLVFF (SEQ ID NO: 143), QKLVF (SEQ ID NO: 144), QKLV (SEQ ID NO: 145), QKL (SEQ ID NO: 146), KLVFFAEDVG (SEQ ID NO: 147), KLVFFAEDV (SEQ ID NO: 148), KLVFFAED (SEQ ID NO: 149), KLVFFAE (SEQ ID NO: 150), KLVFFA (SEQ ID NO: 151), KLVFF (SEQ ID NO: 152), KLVF (SEQ ID NO: 153), KLV (SEQ ID NO: 154), LVFFAEDVG (SEQ ID NO: 155), LVFFAEDV (SEQ ID NO: 156), LVFFAED (SEQ ID NO: 157), LVFFAE (SEQ ID NO: 158), LVFFA (SEQ ID NO: 159), LVFF (SEQ ID NO: 160), LVF (SEQ ID NO: 161), VFFAEDVG (SEQ ID NO: 162), VFFAEDV (SEQ ID NO: 163), VFFAED (SEQ ID NO: 164), VFFAE (SEQ ID NO: 165), VFFA (SEQ ID NO: 166), VFF (SEQ ID NO: 167), FFAEDVG (SEQ ID NO: 168), FFAEDV (SEQ ID NO: 169), FFAED (SEQ ID NO: 170), FFAE (SEQ ID NO: 171), FFA (SEQ ID NO: 172), FAEDVG (SEQ ID NO: 173), FAEDV (SEQ ID NO: 174), FAED (SEQ ID NO: 175), FAE (SEQ ID NO: 176); and (b) the second peptide sequence comprises an amino acid sequence selected from the group consisting of: VDPDNEAYEM (SEQ ID NO: 39), VDPDNEAYE (SEQ ID NO: 40), VDPDNEAY (SEQ ID NO: 41), VDPDNEA (SEQ ID NO: 42), VDPDNE (SEQ ID NO: 43), VDPDN (SEQ ID NO: 44), VDPD (SEQ ID NO: 45), VDP (SEQ ID NO: 46), DPDNEAYEM (SEQ ID NO: 47), DPDNEAYE (SEQ ID NO: 48), DPDNEAY (SEQ ID NO: 49), DPDNEA (SEQ ID NO: 50), DPDNE (SEQ ID NO: 51), DPDN (SEQ ID NO: 52), DPD (SEQ ID NO: 53), PDNEAYEM (SEQ ID NO: 54), PDNEAYE (SEQ ID NO: 55), PDNEAY (SEQ ID NO: 56), PDNEA (SEQ ID NO: 57), PDNE (SEQ ID NO: 58), PDN (SEQ ID NO: 59), DNEAYEM (SEQ ID NO: 60), DNEAYE (SEQ ID NO: 61), DNEAY (SEQ ID NO: 62), DNEA (SEQ ID NO: 63), DNE (SEQ ID NO: 64), NEAYEM (SEQ ID NO: 65), NEAYE (SEQ ID NO: 66), NEAY (SEQ ID NO: 67), NEA (SEQ ID NO: 68), EAYEM (SEQ ID NO: 69), EAYE (SEQ ID NO: 70), EAY (SEQ ID NO: 71), AYEM (SEQ ID NO: 72), AYE (SEQ ID NO: 73), YEM (SEQ ID NO: 74), ATGFVKKDQL (SEQ ID NO:75), ATGFVKKDQ (SEQ ID NO: 76), ATGFVKKD (SEQ ID NO: 77), ATGFVKK (SEQ ID NO:78), ATGFVK (SEQ ID NO:79), ATGFV (SEQ ID NO:80), ATGF (SEQ ID NO:81), ATG (SEQ ID NO: 82), TGFVKKDQL (SEQ ID NO: 83), TGFVKKDQ (SEQ ID NO: 84), TGFVKKD (SEQ ID NO: 85), TGFVKK (SEQ ID NO: 86), TGFVK (SEQ ID NO:87), TGFV (SEQ ID NO: 88), TGF (SEQ ID NO:89), GFVKKDQL (SEQ ID NO: 90), GFVKKDQ (SEQ ID NO: 91), GFVKKD (SEQ ID NO: 92), GFVKK (SEQ ID NO: 93), GFVK (SEQ ID NO: 94), GFV (SEQ ID NO: 95), FVKKDQL (SEQ ID NO: 96), FVKKDQ (SEQ ID NO: 97), FVKKD (SEQ ID NO: 98) FVKK (SEQ ID NO: 99), FVK (SEQ ID NO: 100), VKKDQL (SEQ ID NO: 101), VKKDQ (SEQ ID NO: 102), VKKD (SEQ ID NO: 103), VKK (SEQ ID NO: 104), KKDQL (SEQ ID NO: 105), KKDQ (SEQ ID NO: 106), KKD (SEQ ID NO: 107), KDQL (SEQ ID NO: 108), and KDQ (SEQ ID NO: 109); wherein each of the first peptide sequence and the second peptide sequence optionally includes a C-terminal cysteine. The immunotherapy composition of any one of claims 35 to 36, wherein at least one of the first peptide and the second peptide further comprises a C-terminal portion of the polypeptide or an N-terminal portion of the polypeptide linked to a carrier the linker. The immunotherapy composition of claim 37, wherein the linker comprises an amino acid sequence selected from the group consisting of: GG, GGG, AA, AAA, KK, KKK, SS, SSS, GAGA (SEQ ID NO: 114 ), AGAG (SEQ ID NO: 115) and KGKG (SEQ ID NO: 116). The immunotherapy composition of claim 38, wherein the linker to the carrier optionally comprises a C-terminal cysteine (C). The immunotherapy composition of any one of claims 38 to 39, wherein the carrier comprises serum albumin, immunoglobulin molecules, thyroglobulin, ovalbumin, tetanus toxoid (TT), diphtheria toxoid (DT), Genetically modified cross-reacting substance (CRM) of diphtheria toxin, CRM197, meningococcal outer membrane protein complex (OMPC) and Haemophilus influenzae protein D (HiD), rEPA (Pseudomonas aeruginosa exotoxin A), KLH ( keyhole hemocyanin) and flagellin. The immunotherapy composition of claim 40, wherein the carrier is CRM197. The immunotherapy composition of claim 40, wherein the carrier is diphtheria toxoid. The immunotherapy composition of any one of claims 35 to 42, further comprising at least one pharmaceutically acceptable diluent. The immunotherapy composition of any one of claims 35 to 43, further comprising a multiple antigen presentation system (MAP). The immunotherapy composition of claim 44, wherein the MAP comprises a Lys-based dendritic structure, a helper T cell epitope, an immunostimulatory lipophilic moiety, a cell penetrating peptide, free radical induced polymerization, presented as an antigen One or more of self-assembled nanoparticles and gold nanoparticles of the platform. A pharmaceutical composition comprising the immunotherapy composition of any one of claims 35 to 45 and at least one adjuvant. The pharmaceutical composition of claim 46, wherein the adjuvant is selected from the group consisting of: aluminum hydroxide, aluminum phosphate, aluminum sulfate, 3-de-O-acylated monophosphoryl lipid A (MPL), QS -21, TQL1055, QS-18, QS17, QS-7, complete Freund's adjuvant (CFA), incomplete Freund's adjuvant (IFA), oil-in-water emulsions (such as squalene or peanut oil), CpG, poly Glutamic acid, polylysine, AddaVax™, MF59® and combinations thereof. The pharmaceutical composition of claim 47, wherein the adjuvant is QS-21 or TQL1055. The pharmaceutical composition of claim 47, wherein the adjuvant is MPL. The pharmaceutical composition of claim 47, wherein the adjuvant is a combination of MPL and QS-21, or a combination of MPL and TQL1055. A nucleic acid comprising a nucleic acid sequence encoding the polypeptide of any one of claims 1 to 21 in the immunotherapy composition of claims 35 to 39. A nucleic acid immunotherapy composition comprising the nucleic acid of claim 51 and at least one adjuvant. A method of treating or achieving prevention of Alzheimer's disease in an individual comprising administering to the individual an immunotherapy composition as claimed in any one of 22 to 25 and 35 to 45 or as claimed The pharmaceutical formulation of any of 26-34 and 46-50. A method of inhibiting or reducing the aggregation of at least one of Aβ and α-synuclein in an individual with Alzheimer's disease or at risk of developing Alzheimer's disease, comprising administering to the individual The immunotherapy composition of any of claims 22-25 and 35-45 or the pharmaceutical formulation of any of claims 26-34 and 46-50 is administered. A method of treating or effecting prevention of Alzheimer's disease in an individual comprising administering to the individual a nucleic acid immunotherapy composition as claimed in claim 52. A method of inhibiting or reducing the aggregation of at least one of Aβ and α-synuclein in an individual with Alzheimer's disease or at risk of developing Alzheimer's disease, comprising administering to the individual The nucleic acid immunotherapy composition of claim 52 is administered. The method of any one of claims 53 to 56, further comprising at least two, at least three, at least four, at least five, or at least six repeated administrations. The method of claim 57, further comprising repeating the administering at intervals of about 21 to about 28 days. A method of inducing an immune response in an animal, comprising administering to the animal any one of the following: the polypeptides of claims 1 to 21, the polypeptides of claims 22 to 25, and 35 to 45 in a regimen effective to generate an immune response The immunotherapy composition of claims 26 to 34 and 46 to 50, or the nucleic acid immunotherapy composition of claim 52, the immune response comprising specific binding to Aβ, α-synuclein or Aβ and antibodies to both α-synuclein. The method of claim 59, wherein the immune response comprises an antibody that specifically binds to A[beta] and an antibody that specifically binds to [alpha]-synuclein. The method of any one of claims 59 to 60, wherein the inducing an immune response comprises an antibody that specifically binds to the N-terminal region of Aβ and/or the C-terminal region of α-synuclein. An immunization kit comprising the immunotherapy composition of any one of claims 22 to 25 and 35 to 45. The kit of claim 62, further comprising an adjuvant. The kit of claim 63, wherein the immunotherapy composition is in a first container and the adjuvant is in a second container. A kit comprising the nucleic acid immunotherapy composition of claim 52. The kit of claim 65, further comprising an adjuvant. The kit of claim 66, wherein the nucleic acid is in a first container and the adjuvant is in a second container.

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